Large Animal Review 4-2023

Large Animal Review

ORIGINAL ARTICLES

BOVINE

ISSN: 1124-4593

ANIMAL REVIEW is ranked in Citation Index (SciSearch®) Journal Citation Reports/Science Edition and CAB ABSTRACTS

• Effect of a blend of essential oils, bioflavonoids and tannins on production performance, health, immune functionality, and antioxidant status in fattening beef cattle

CASE REPORTS

BOVINE

• Outbreaks of lethal cyanogenic glycosides poisonings of cattle after ingestion of Sorghum ssp. grown under drought conditions in August 2022 in Piedmont (NorthWestern Italy)

• Treatment of an atresia ani et recti and urogenital agenesis in a calf

• Comparison of the effect of two different joint lavage techniques on survival rate in calves with septic arthritis: 248 cases

• Systemic embolic hepatitis and pneumonia caused by subacute ruminal acidosis (SARA) in Hanwoo (Korean indigenous) cattle: A case report

EQUINE

• Pre-maxilla fracture repair using interfragmentary stainless steel wire in a foal

LARGE

ITALIANA VETERINARI PER ANIMALI DA REDDITO

1 University of Milan, Department of Veterinary Medicine and Animal Science, Via dell’Università 6, Lodi, 26900, Italy;

2 Doctor of Veterinary Medicine;

3 PhD - Animal Nutrition and Food Safety

SUMMARY

The aim of the study was to evaluate the effect of a blend of essential oils, bioflavonoids and tannins on production performance, health, immune response, and antioxidant status in fattening beef cattle under field conditions.

A total of 210 newly arrived male Charolaise bulls were assigned to two study groups: i) Control (417.84 ± 19.63 kg live weight), basal diet; ii) Treatment (416.37 ± 18.56 kg live weight), basal diet integrated with 5 g/head/d of the blend of essential oils, bioflavonoids, and tannins. Growth performances, dry matter intake (DMI), feed conversion rate (FCR), carcass characteristics and health were evaluated during the entire fattening period (182 days). The immune response and the oxidative status were evaluated at the arrival (d0), and after 45 days (d45) by the titration of antibodies against BHV-1, serum bactericidal activity, γ-interferon levels, free oxygen radical metabolites (ROM’s), and antioxidant activity (OXY).

Growth performances and production efficiency resulted to be significantly improved by the treatment. Indeed, both weight at d102 and d182 were higher in the Treatment group (respectively 579.81 and 720.86 kg vs 572.69 and 709.82 kg in the Control) (P≤0.05).

As a result, also the total average daily gain (ADG0-182) (1.67 vs 1.60 kg/head/d) (P≤0.001) and the ADG between d0 and d102 (ADG0102) (1.60 vs 1.52 kg/head/d) (P≤0.05) were improved. The DMI was lower in the Treatment group (10.59 vs 11.18 kg/head/d) (P≤0.001), leading to a significant improvement in the FCR (6.37 vs 7.04) (P≤0.001). Carcass weight was higher in the Treatment group (425.87 vs 418.31 kg in the Control group) (P≤0.001), while carcass characteristics weren’t affected by the treatment. The incidence of bovine respiratory disease (BRD) was significantly reduced by the treatment (17.14 vs 28.57 % in the Control group) (P≤0.05). No differences were foundin the incidence of lameness. The Treatment has led to an increase in the serum bactericidal activity (92.00 vs 80.40 % in the Control group) (P≤0.05) and to a better antioxidant status (365.68 vs 290.58 µmol HClO/ml in the Control group) (P≤0.001) at d45. No differences were found in terms of antibodies titration and ROM’s levels. In conclusion, the supplementation of fattening beef cattle diet with a blend of essential oils, bioflavonoids and tannins, improved growth performance, feed efficiency and health status thanks to a better feed efficiency and immune functionality.

KEY WORDS

Beef cattle, efficiency, natural products, animal health, immunity, welfare.

INTRODUCTION

The zootechnical sector will face many challenges in the next years. The overall increase in the world population, prospected to double by 2050, together with a generalized improvement in the economic and social status, will lead to a steady growth in the demand of animal derived foods [1-5]. Moreover, those developments of the human society can increase pressure on some resources that are already scarce, such as land and water, reducing their availability for animal production [5]. Furthermore, there is a constant increase in the consumers’ awareness about animal welfare and antibiotic use in farming animals, also due to the increased spread and frequency of dis-

Corresponding

Author:

Silvia Grossi(silvia.grossi@unimi.it)

ease caused by antimicrobial resistant bacteria. Indeed, an excessive and incorrect use of antibiotics at the farm level, frequently driven by poor welfare conditions, is one of the possible triggers for the development of those resistant strains, together with both an incorrect and excessive use in human medicine and an improper management of wastes and wastewaters [6]. Moreover, the exacerbating environmental crisis has increased the pressure about the role of zootechny on climate change and other environmental related topics, such as soil and water pollution and water and agricultural land scarcity [7-8].

The need and urgency to address, in a short time, all those targets are highlighted also by the newest national and European rules on animal welfare and antimicrobials use in animal farming, and by the new allocation of the PAC subsidies in relation to the achievement of specific environmental targets [9].

Between all the food producing animals, beef cattle are among the most criticized. Beef cattle farming is responsible of about 35% of the total livestock farming methane emissions [10], and

C.A. SGOIFO ROSSI1, S. GROSSI*1, R. COMPIANI2, G. BALDI3

Effect of a blend of essential oils, bioflavonoids and tannins on production performance, health, immune functionality, and antioxidant status in fattening beef cattle

moreover beef cattle are the less efficient in converting the feeds used into final edible products, compared to other food producing species [11-13]. Indeed, beef cattle are characterized, on average, by a feed conversion rate of 6 to 10 per kg of protein produced, while the ones of pigs (2.7 to 5) and chickens (1.7 to 2) are significantly lower [11-13]. In terms of health problems and antimicrobial use, intensive beef cattle farming is in third position in terms of milligrams per population correction units (PCU) at a global level, behind swine and poultry (172 mg/PCU, 148 mg/PCU, and 45 mg/PCU, respectively) [14]. However, there are still stages, such as the arrival period, that can represent a treat for animal health and welfare, thus frequently requiring a higher use of antimicrobials [15,16]. Indeed, newly received cattle are exposed to several stressors, such as weaning, long-distance transport, mixing, feed and water restrictions, and adaptation to new environmental and feed conditions [15]. All these factors contribute to pathogen colonization and proliferation as well as to a stress-related reduced immune functionality, that can lead to a higher incidence of diseases, especially bovine respiratory disease (BRD) [15-17]. Consequently, higher quantities of antibiotics are often used in this stage, both to treat sick subjects but also to prevent the spread of the problem [15].

New strategies must be designed, tested, and implemented to maximise beef cattle health and welfare, with a consequent reflection on production efficiency and environmental sustainability, due to the strong correlations between all those parameters. As an example, the onset of BRD in the first days after the arrival can cause a strong reduction in the growth performances with a reflection on the entire fattening period [18]. This led to both a waste of feed resources, often in competition with human nutrition or already scarce, such as water, as well as to a lower quantity of final products per animal. Basically, if efficiency and health are not maximized more animals are required to produce the same amounts of final products, leading to a both a higher production of methane and to an increased need of feed products, increasing the demand of scarce and critical resources, such as water and agricultural land. In these directions, animal nutrition can have a proactive role, improving production performance, feed efficiency, and animal welfare, and reducing the risk of pathologies, improving consequently, the environmental impact. Indeed, besides being functional to fulfil all the basic nutritional requirements, nutrition can be a helpful tool to modulate animal health and efficiency, and consequently the overall sustainability levels [19]. Firstly, through nutrition is possible to maintain and even enhance rumen functionality and efficiency [20]. A correct and balanced diet is the main tool to avoid imbalances and sudden changes in the ruminal environment, that can impair its activity, leading thus to the onset of digestive disorders, such as acidosis [18,20]. Those diseases, besides reducing the overall productivity and production efficiency of the animals, are often the door opener and the trigger for other health problems, such as lameness, bloat, and enterotoxaemia [18].

At the same time, nutrition can be a vehicle to administer bioactive compounds to support the rumen as well as the general immune system. Indeed, rumen health and functionality can be safeguard and improved by additives, such as yeasts, organic minerals, and natural extracts, able to influence rumen microflora and the digestive processes [15-19]. Moreover, modulating the ruminal environment and consequently the ruminal microflora through the feed and some bioactive molecules, it

is possible to reduce the overall methane production [19]. At the same time, similar additives can also have a modulatory effect on the general immune system, providing bioactive compounds that can sustain both the antioxidant defences and the innate and adaptive immune response [15-23].

Between natural products, essential oils, bioflavonoids and tannins are showing promising result in the application in ruminant’s nutrition due to their proven ability in modulate the ruminal microflora toward a more efficient environment and methanogenesis reduction [19, 24-27] and in improving health status thanks to their antimicrobial, antioxidant and immunostimulant properties [28].

Those properties are variable between the different natural compounds as well as between the different blend of them. Their effectiveness, and the specific targets for which they are used, varies depending on their molecular characteristics, their composition, and their combinations. Also, the quantities administered as well as the form and duration of the administration and other factors related to the nutritional and overall management of the animals, can affect their functionalities. Those aspects lead to different possibility of application of the different natural products, as well as to potentially controversial results.

A combination of coriander, geraniol oils and bioflavonoids and tannins has been already tested in dairy cows, reporting positive effects on production performances such as milk yield, feed conversion efficiency and on in vivo diet digestibility [19]. This combination of natural products has still to be evaluated in terms of effect on the immune systems as well as specifically on beef cattle. However, there are some evidence that single components of this mixture can positively affect beef cattle growth as well as the immune functionality and antioxidant defences in beef and dairy cows [29-33].

On the base of these evidence, the aim of the present study was to evaluate the effect of the inclusion of a blend of coriander, eugenol, geraniol oils and bioflavonoids and tannins (Anavrin, Vetos Europe Sagl - Cadenazzo TI - Switzerland) in the diets of fattening beef cattle on production efficiency, health, immune response, and antioxidant status under field condition.

MATERIALS AND METHODS

The procedures relating to animals were carried out in compliance with the directive of the Council of the European Communities (2010/63 / EU), implemented by the Italian Ministry of Health (Legislative Decree 26, March 4, 2014).

This trial was set up to evaluate the efficacy of a blend of essential oils (EO), mainly from cloves (Syzygium aromaticum), coriander seed (Coriandrum sativum), and geranium (Pelargonium cucullatum), tannins (CT) from chestnuts (Castanea sativa) and bioflavonoids (BF) from olives (Olea europea) (relative concentrations of the active principles were: EO:CT:BF = 1:2.5:0.1) (Anavrin, Vetos Europe SAGL, - Cadenazzo TI, Switzerland), on beef cattle productivity, health, immune response and antioxidant status.

Animals, housing, and trial groups

The study took place in an intensive beef fattening unit, located in northern Italy (via Viola 6, 37050 Roverchiaretta, (VR), Italy), that well-represent the typical Italian intensive beef cattle fattening farms.

A total of 210 Charolaise bulls , imported from France, were enrolled at the arrival (d0), after being individually weighed and evaluated for body conformation using a 5-point scale (1: profiles straight and poor muscle development; 2: profiles between whole straight to low convex and medium muscle development; 3: profiles low convex and good muscle development; 4: profiles on the whole convex and very good muscles development; 5: all profiles convex and exceptional muscles development) [34].

The animals were then grouped by weight and conformation, and randomly assigned to two balanced groups, differing only for the inclusion or not in the diet of the blend of natural products: i) Control (105 heads, 417.84 ± 19.63 kg live weight), basal diet; ii) Treatment (105 heads, 416.37 ± 18.56 kg live weight), basal diet integrated with 5 g/head/d of that blend.

The bulls were housed on slatted floor in a close barn, in 30 pens with 7 animals each (3.5 m2 each).

The trial lasted for the entire fattening period (182 days).

Nutritional management

The two study groups were fed following the same nutritional plan (Table 1), characterized by two different formulations (arrival and fattening), studied to meet the specific growth needs in those different phases, in agreement with the Nutrient Requirement Council [35]. The feed was administered in the form of total mixed ration (TMR).

Two separate TMRs, equal in terms of nutritional values but differing for the inclusion or not of the treatment, were produced daily. Indeed, the pool of natural products was included directly in the mineral mix used in the TMR administered to the Treatment group, to optimize the mixing and to guarantee its inclusion at 0.5-1 g/kg d.m. (dry matter) of feed. The same amount of wheat powder was added in the mineral mix of the Control group as a placebo.

The TMRs were administered ad libitum for the entire fattening period and delivered once a day in the morning by a feed mixer wagon, provided with electronic scale to weigh the inclusion of each ingredient and the amount of the TMR unloaded. Water was available ad libitum

Parameters recorded

Growth and slaughtering performances

Individual body weight was recorded before morning feeding at three timepoints, enrolment day (d0), day 102 (d102) and before slaughter (d182). The individual average daily gain (ADG) was then calculated for each period, from d0 to d102, from d102 to d182 and from d0 to d182, using the following formula:

ADG = Weightf - Weighti days i - f

Where:

ADG= average daily gain (kg/head/day)

Weight f= final weight of each period

Weight i= initial weight of each period

Days i-f= days between the start and the end of each period

The daily feed intake of each pen in the two groups was evaluated weekly by weighing the TMR administered and the residue in the manger 24 h later. Then the feed intake was corrected for the dry matter level of the diet, to obtain the dry matter intake (DMI), The FCR was then calculated, comparing the average DMI intake of each pen from d0 to d182, with the ADG

of the corresponding pen in the same period. Data about cold carcass weight, dressing percentage, carcass conformation and fattening (SEUROP) scores were collected for all animals at the slaughterhouse. The cold carcass weight was recorded after 24h of chilling at a temperature of 0°C to 4°C. The dressing percentage was obtained comparing the final live weight with the cold carcass weight. Carcass conformation and fattening scores were assessed by an expert judge following the EU legislation (Council Regulation EEC n. 1026/91, 22 April 1991) [36] , using the SEUROP classification method, with a conformation scale ranging from S to P (S-superior: all profiles extremely convex, exceptional muscle development, double-muscled conformation; E-excellent: all profiles convex to super-convex, exceptional muscle development; U-very good: profiles on the whole convex, very good muscle development; R-good: profiles on the whole straight, good muscle development; O-pretty good: profiles straight to concave, medium muscle development; P-poor: all profiles concave to very concave, poor muscle development), and a fattening scale ranging from 1 to 5 (1low: none up to low fat cover; 2- slight: slight fat cover, flesh visible almost everywhere; 3-medium important: flesh, with the exception of the round and shoulder, almost every-where covered by fat, slight fat deposits in the thoracic cavity; 4-high: flesh covered by fat, round and shoulder still partly visible, medium fat deposits in the thoracic cavity; 5-very high: carcass well covered by fat, heavy fat deposits in the thoracic cavity).

Table 1 - Feeding plan: composition and predicted nutritional value of the two diets used during the entire fattening period, as calculated by the rationing software (Plurimix).

1CP= crude protein; 2d.m.= dry matter; 3NDF= neutral detergent fiber; 4ADF= acid detergent fiber; 5ADL= acid detergent lignin; 6Ca= calcium; 7P= phosphorus

of a blend of essential oils, bioflavonoids and tannins on production performance

Health status, immune response, and antioxidant status

The individual health status was checked twice a day by the veterinary and qualified animal health care staff of the farm during the entire fattening period. Any cases of morbidity and mortality were recorded, as well as the number of animals that needed to be moved to the infirmary pen, together with the motivation, with specific attention on the incidence of bovine respiratory disease (BRD) and lameness. Sick animals were treated according to the procedures, medications, and sanitary protocols adopted by the farm veterinary staff.

Blood samples were taken at the arrival (d0) and at d45 on a subsample of animals (5 heads in each group) to evaluate some serum indicators of immune response and antioxidant status. Blood samples were collected by jugular venepuncture into 10mL EDTA tubes and 10-mL no additive tubes (Venoject®, Terumo Europe N.V., Leuven, Belgium) and immediately placed on ice. Then, the serum was extracted by centrifugation at 3000 g for 10 min at 4 °C and stored at - 20 °C. The immune response was evaluated in terms of both specific and nonspecific immunity and in terms of oxidative status.

The titration of BHV-1 vaccination antibodies with a BHV-1 serum neutralization test was evaluated as an indicator of specific immunity modulation. The BHV-1 serum neutralization test was performed according to OIE (2012).

Non-specific immunity was evaluated with the analysis of the serum bactericidal activity and γ-interferon.

The oxidative status was evaluated through the analysis of both the amount of free oxygen radicals’ metabolites (test ROM’s) and of the antioxidant activity (test OXY).

The amount of free oxygen radicals in plasma samples was determined using the d-ROMs test (Diacron, Grosseto, Italy), which determines hydroperoxides (the breakdown products of lipids, as well as of other organic substrates, generated by the oxidative attack of ROS) through their reaction with the chromogen N,N-diethyl paraphenylenediamine. Results are expressed in arbitrary Carratelli Units (U CARR), where 1 (U CARR) is equivalent to the oxidizing power of 0.08 mg H2O2/dL.

The antioxidant activity was evaluated through the OXY-adsorbent test. This method measures the ability of a sample to oppose a massive oxidant attack, induced in vitro, by a hypochlorous acid solution and the results are expressed in terms of moles HClO/mL (hypochlorous acid).

Statistical analysis

The statistical analysis of the data was conducted using the SAS

statistical software (SAS 9.4, SAS Cary NC).

For the evaluation of the growth performances the pen was used as an experimental unit. These data were analysed using a mixed model (PROC MIXED) which considered the fixed effect of the treatment and the time of detection and the random effect of the pen.

The single subject was instead used as a reference unit for evaluating the characteristics of the carcass using a mixed model (PROC MIXED) which considered the fixed effect of the treatment. Also, the single subject was used as a reference unit for the evaluation of the parameters related to the immune functionality, using a mixed model (PROC MIXED) which considered the fixed effect of the treatment and the time of detection.

For non-continuous variables, such as SEUROP classification, fattening status and health status, the difference in frequency distribution within the classes was evaluated by applying a chisquared test (PROC FREQ).

The difference was considered significant for P ≤0.05, while a tendency was set up at P<0.1.

RESULTS AND DISCUSSION

Growth and slaughtering performances

Data related to growth performance and production efficiency are reported in Table 2. The treatment has led to an improvement ADG, both considering the first period of the trial (ADG0-102) (1.60 vs 1.52 kg/head/d in the Control group) (P ≤0.001) and the overall fattening period (ADG0-182) (1.67 vs 1.60 in the Control group) (P ≤0.05). Specifically, this difference was equal to 80 g/head/d during the first 102 days following arrival and 70 g/head/d considering the entire rearing period. On average, at the end of the fattening cycle, there was a 4.4% of increase in terms of ADG in the Treatment group.

Those improvements in the daily gain have led to higher weights in treated animals, both at d102 (579.81 vs 572.69 kg in the Control group) (P ≤0.05) and at the end of the fattening period (720.86 vs 709.82 kg/head/d in the Control group) (P ≤0.05). Moreover, a lower DMI was reported in the Treatment group (10.59 vs 11.18 kg in the Control group) (P ≤0.001) during the entire fattening period. This reduction, together with the improvement in the growth performances has led to an optimization of the feed efficiency (FCR: 6.37 vs 7.04 in the Control group) (P ≤0.001), resulting in an overall 9.52% improvement in FCR in treated animals.

This increase in production efficiency can be explained by a pos-

Table 2 - Growth performance in the two study groups.

itive and stimulating action of the blend of natural compounds on the ruminal microflora, resulting in a more efficient ruminal environment. This aspect can improve the efficiency of utilisation of the feeds by the ruminal microflora, leading to a higher percentage of volatile fatty acids, especially of propionate, produced in the rumen [19,37,38]. Sgoifo Rossi et al. [19], in a study done in dairy cows, have found a significant improvement of the in vivo diet digestibility of starch and cellulose in cows treated with the same blend of natural products, explained by a more viable and functional ruminal microflora. Indeed, several studies based on similar natural compounds have shown an increase in the main ruminal populations involved in both structural and non-structural carbohydrates degradation, such as Ruminococcaceae and Propionic acid bacteria [39,40], with an improvement in propionate production and the acetate:propionate ratio [40]. As an example, Klop et al. [40], in an in vitro incubation test reported a higher proportion of propionate in the ruminal fluid of donor cows, that received a blend of similar natural compounds, compared to control cows that did not receive the integration. Moreover, different bibliographical studies, done both in vitro and in vivo, have reported a potential inhibitory activity of some of the natural compounds present in the blend used in the present study on ruminal methanogenic bacteria, highlighting how this can increase the bioavailability of substrates for other microbial populations, leading to a greater production of volatile fatty acids [19,27,39-42]. Consequently, those increased quantities of volatile fatty acids available for the animal can explain the increase in the overall growth and productive parameters as well as the overall reduction in the feed intake, since a higher amount of energy is already available at the ruminal level. Because of the better rumen functionality and diet digestibility, Sgoifo Rossi et al. [19] found a 3.8% increase in daily milk, together with a 7.2% of increase in terms of FCR production in dairy cows fed with the same blend of natural products. Those results are also confirmed in other studies, that also highlighted that shorter administration periods can affect the results, with lower efficacy in shorter studies [27]. Those results obtained in dairy cows are in line with the finding of the present study, even if it is necessary to consider the differences between dairy and beef cattle, as well as the differences in their management, especially considering the nutritional plans.

Nowadays, there are not studies done in beef cattle with the same blend of essential oils, that evaluate comprehensively the effectiveness of this specific combination. However, there are some evidences that its components, alone or in combination with other natural products, can positive-

ly affect growth performance and feed efficiency in beef cattle. Alves de Souza et al. [30] found an increase in the ADG (+0.240 g/head/d) in fattening beef heifers that received a mixture of clove and different natural products compared to control heifer without any integration [30]. Conversely, in that study the DMI resulted to be increase, partially explaining the increase in the growth performance [30]. Also, Compiani et al. [28], found a significant increase in the ADG and overall growth performances in beef cattle treated with a pool of different natural compounds, also containing eugenol, one of the main bioactive compounds present in clove. Moreover, different studies have reported some positive effects of tannins on growth performance of beef cattle fed with corn-based diets [30-33]. Indeed, even if high concentrations of tannins may be toxic, reducing voluntary feed intake and nutrient digestibility, however at low to moderate concentrations, tannin supplementation may shift site of protein degradation increasing metabolizable amino acid flow to the small intestine [43-44]. This tannin effect may explain the improvements observed in performance of feedlot cattle [32-33]. Conversely, Tabke et al. [45] did not found any improvement in terms of growth performance in beef steers that received an integration of tannic acid. Conversely, a reduction in the apparent total tract digestibility of starch and protein was found in treated animals [45].

Data related to slaughtering performances are reported in Table 3. Carcass weight resulted to be higher in the Treatment group (425.87 vs 418.31 kg) (P ≤0.001), as a reflection of the higher live weight achieved at the end of the fattening period. No significant differences were found in terms of dressing percentage, conformation, and fattening score between groups. No specific studies are available in the bibliography that evaluate the effect of the same blend of natural products on carcass quality and characteristics. However, there are some studies that tested the effect of some of its components, alone or in combination with other natural products, on carcass characteristics. As in the present study, Mottin et al. [46] have found no effect of the inclusion of a blend of natural products, containing clove and other compounds, on the main parameters related to the carcass evaluation, such as carcass yield, subcutaneous fat thickness, muscle area and marbling. Also, Tabke et al. [45] did not found any effects of the inclusion of tannic acids on carcass yield and quality, expressed as quality grade and yield grade.

Health status, immune functionality, and antioxidant status

Data related to the health status, recorded during the entire trial, are reported in Table 4. No cases of mortality were record-

Table 3 - Carcass characteristics and slaughtering performances in the two study groups.

ed. Moreover, no animals in both groups needed to be moved to the hospital pens due to severe health issues that required isolation and specific care.

In terms of pathological issues, the treatment has led to an overall reduction of the incidence of BRD (28.57% vs 17.14%) (P ≤0.05), because of a tendency toward a reduction of both the incidence of first cases (13.33 vs 19.05 in the Control group) (P=0.08) and in the number of relapses (3.81 vs 8.59 in the Control group) (P=0.08).

Conversely, no differences were found in terms of lameness incidence, that was similar in the two groups and in line with a value corresponding to a low incidence [47].

The positive effects on health registered during the overall fattening period in the Treatment group may be justified by a better non-specific immune reactivity and antioxidant status, as highlighted by the immunological results reported in Table 5, especially during the arrival phase, that is the most critical period of fattening cycle, and the one in which most of the health issues are registered.

Specifically, the treatment has led to a higher serum bactericidal activity at d45 (92.00 vs 80.40 in the Control group) (P≤0.05), resulting significantly above the 90% threshold, a value considered to be the limit for healthy cattle [48]. Moreover, the treatment has led to an improvement in the antioxidant defences, as highlighted by the significantly higher results obtained in terms of antioxidant activity (365.68 vs 290.58 µmol

HClO/ml in the Control group) (P≤0.05). No difference was found in specific immune response to BHV-1 vaccine, and on γ-interferon and ROMs values.

Even if, also in terms of the evaluation of health and immune parameters, no studies that tested the same combination of natural products are available in bibliography, there are still some evidences related to some of the single compounds. Compiani et al. [28] found that the inclusion of a mixture on natural compounds, also containing eugenol, has led to a significant increase in the bactericidal activity during the first period of the fattening cycle [28]. Indeed, different studies have reported a strong bactericidal activity of clove against different pathogens, including some related to BRD, such as Pasteurella multocida [48-49].

Also, Santillo et al. [29] found a positive effect of feeding tannins to lactating diary cows on the main parameters related to the antioxidant status. As in the present study, in which the highest levels reached in the OXY test highlight a better efficiency of the antioxidant defences, Santillo et al. [29] found an increased biological antioxidant potential in the treated animals. The inclusion of tannins in the dairy cows’ diet was also able to reduce the presence of ROMs in the serum of treated animals. Due to the low percentage of direct absorption of tannins, the effect on the antioxidant status and antioxidant defences can be explained by a local activity in the gastrointestinal tract, that can lead to an increase in the expression and activity of some endogenous antioxidant enzymes [50-51].

Table 5 - Immunological parameters in the two study groups evaluated at d0 and d45.

Table 4 - Health status in the two study groups during the entire fattening period.

CONCLUSIONS

The results of the present study highlight that inclusion of a blend of essential oils, bioflavonoids, and tannins in the diet of fattening beef cattle can improve the overall production efficiency. Indeed, this blend of natural product can exert a proactive role on both the ruminal efficiency and health, modulating the fermentation kinetics and increasing thus the synthesis of volatile fatty acids used for energy purpose. Moreover, the inclusion of that blend of natural products can improve the immune functionality and in the antioxidant defenders, factors that may have influenced the health status of the treated animals, leading to a lower incidence of bovine respiratory disease. The combination of those aspects increase the production efficiency of fattening beef cattle, also reducing the need of antimicrobials due to an improved welfare status. On average, the inclusion of this blend of natural products can led to a more sustainable way to raise beef cattle.

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Piedmont (North-Western Italy)

STEFANO GIANTIN1, ALBERICO FRANZIN1*, GIOVANNI TOPI2, GIORGIO FEDRIZZI3,

CARLO NEBBIA4

1 Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta - Cuneo, Italy

2 Veterinario libero professionista - Marene (CN), Italy

3 Laboratorio Nazionale di Riferimento per le Tossine Vegetali negli alimenti, Istituto Zooprofilattico Sperimentale di Lombardia ed Emilia-Romagna - Bologna, Italy

4 Dipartimento di Scienze Veterinarie, Università degli studi di Torino - Italy

SUMMARY

The following case report aims to present an outbreak of cattle poisoning recorded in Piedmont (Northwestern Italy) during the summer 2022. In the first half of August several accidents occurred in five different farms located in the provinces of Cuneo, Asti and Biella. The first and most severe outbreak was reported in a grazing field located in Sommariva del Bosco (Cuneo) and involved a herd of Piedmontese heifers which ingested a hybrid cultivar of Sorghum bicolor x S. sudanense resulting in the death of 50 animals. Sixteen additional deaths were recorded in the fo llowing days in the remaining four farms. In all the latter cas es, poisoned cows had been fed with the widespread weed S. halepense. Interestingly, farmers complained that no such events have ever been recorded in all previous years, feeding cattle with or grazing on sorghum being a common practice. Shortly after exposure, the affected bovines showed recumbency, sialorrhea, tachypnea, air hunger and muscle tremors followed by acute depression and death, pointing thereby to cyanide poisoning. Most of animals received an oral or intravenous treatment with the antidote sodium thiosulfate, albeit this compound is not typically referred to as the sole remedy against cyanide toxicosis. All treated animalsshowed a complete recovery. Carcasses of dead animals were subjected to necropsy and sorghum samples were collected in each farm/field to determine the content of the cyanogenic glycoside dhurrin by LC-MS/MS. As a result, dhurrin concentrations up to 10,000 mg/kg were detected in four plant samples collected from the respective involved farms. The anamnesis, the rapid onset, the clinical picture, the remarkable efficacy of the antidote and the results from necropsies strongly indicated cyanide intoxication. This diagnosis was further supported by the high dhurrin concentrations measured in most sorghum samples, which are consistent with the extreme drought conditions that affected the area including the mentioned farms. In that area, the Regional Agency for Environmental Protection (ARPA) reported several days with summer tropical diurnal and nocturnal temperatures as never experienced before. Therefore, caution is needed when feeding cattle with green chops or while grazing, with a special care in case of potential drought stress suffered by Sorghum plants.

KEY WORDS

Cyanide; Cattle poisoning; Dhurrin; Sorghum; Drought.

INTRODUZIONE

Il sorgo (genere Sorghum) viene coltivato per la nutrizione animale sia come foraggio fresco che per la produzione di fieno e insilati. Seppur con caratteristiche bromatologiche meno pregiate ed una minor digeribilità rispetto ad altre foraggere [1], il sorgo, grazie alla tolleranza alla siccità e la resistenza agli agenti infestanti, rappresenta una delle principali alternative al mais quale fonte di alimentazione per i ruminanti nelle zone aride [2]. In varie regioni del nord Italia viene da tempo usata come foraggio fresco anche la specie selvatica S. halepense (L.) Pers, a carattere infestante e comunemente definita sorghetta. Il sorgo, come meccanismo di difesa, produce il glucoside

Corresponding Author: Alberico

Franzin (alberico.franzin@izsto.it)

cianogenetico (S)-(-D-Glucopyranosyloxy)(4-hydroxyphenyl) acetonitrile, durrina [3]. Quando i tessuti della pianta vengono lacerati, come in caso di masticazione da parte di erbivori, il cianuro (-CN) viene liberato dalla durrina. Inoltre, i microrganismi presenti nel rumine dei bovini sono in grado di accelerare ulteriormente l’idrolisi e la seguente liberazione di -CN [4]. Tale molecola, legandosi al Fe+++ della citocromoossidasi mitocondriale, blocca la respirazione cellulare: l’ossigeno (O2) non viene più utilizzato dai tessuti e viene bloccata la produzione di energia immagazzinata sotto forma di ATP [5].

Il sorgo produce naturalmente durrina durante le prime fasi di sviluppo; infatti, si consiglia di non somministrare piante giovani agli animali per via dell’elevata concentrazione del glicoside [6, 7]. I nuovi germogli, le foglie giovani e i ricacci risultano essere le parti più pericolose poiché accumulano maggiori quantità del metabolita [8]. Le piante adulte sono

Outbreaks of lethal cyanogenic glycosides poisonings of cattle after ingestion of Sorghum ssp. grown under drought conditions in August 2022 in

172Outbreaks of lethal cyanogenic glycosides poisonings of cattle after ingestion of Sorghum ssp. grown

generalmente considerate sicure come alimento animale [9] ma, in alcune condizioni ambientali stressanti, possono produrre grandi quantità di durrina. Siccità estreme e prolungate seguite da intense piogge o copiosa irrigazione, forti gelate seguite da periodi più caldi, così come trattamenti con alcuni erbicidi o l’utilizzo di fertilizzanti azotati possono stimolarne un eccessivo accumulo. In genere il processo di fienagione e l’insilamento permettono la dispersione del -CN dai tessuti vegetali, anche se in alcuni casi la concentrazione può rimanere alta, ad esempio se il sorgo si essicca rapidamente ed è conservato in balle di fieno di grandi dimensioni [10].

IL CASO

Di seguito sono descritti cinque casi di intossicazione avvenuti in Piemonte nel mese di agosto 2022, nell’arco di meno di tre settimane, durante un periodo di forte siccità accompagnato da temperature diurne e notturne superiori alle medie stagionali.

A. Il 6 agosto a Sommariva del Bosco (CN) 160 manze di razza prevalentemente piemontese venivano liberate su un campo seminato a S. bicolor x S. sudanense varietà Suzy [11]. I capi, affamati dal digiuno notturno, hanno assunto il foraggio con grande voracità. Trattavasi di un ricaccio verde di altezza 3045 cm, a seguito del primo sfalcio avvenuto il 14 luglio. 80 capi hanno presentato sintomi da intossicazione. 20-30 minuti dopo l’ingestione, 50 manze, quasi tutte gravide, sono decedute. I restanti capi sono stati immediatamente allontanati dal pascolo.

B. L’11 agosto a Moretta (CN) è stata somministrata erba di sfalcio fresca, contenente prevalentemente S. halepense, a un gruppo di 20 vacche di razza mista da latte, di età superiore a 3 anni. Tutti gli individui sono risultati intossicati e 5 di essi sono venuti a morte.

C. L’11 agosto a Bra (CN) un gruppo di 60 vacche di razza piemontese, di età superiore a 3 anni, è stato alimentato con foraggio costituito perlopiù da sorghetta. 36 animali sono stati colpiti da avvelenamento, 6 sono deceduti.

D. Il 12 agosto ad Asti sono deceduti 4 bovini di razza piemontese mentre pascolavano su un campo in cui era presente sorgo selvatico. Causa segnalazione tardiva, non si sono potute registrare la consistenza della mandria e la morbilità dell’avvelenamento.

E. Il 25 agosto a Cossato (BI) una mandria di 45 bovini prevalentemente di razza piemontese ha pascolato in un campo ove cresceva sorghetta. Tutti i capi hanno presentato sintomi di intossicazione, 1 è morto.

SINTOMATOLOGIA

Tutti gli animali intossicati hanno presentato in modo più o meno uniforme i sintomi riportati in Tabella 1.

MATERIALI E METODI

Sono stati effettuati esami necroscopici sugli animali deceduti in ogni azienda colpita a eccezione del caso D. Per ogni episodio si è provveduto al campionamento di almeno 500 g di sorgo fresco prelevato in differenti posizioni del campo e tempestivamente congelato. I campioni sono stati inviati al Laboratorio Nazionale di Riferimento per le Tossine Vegetali negli alimenti dell’Istituto Zooprofilattico Sperimentale di Lombardia ed Emila Romagna, ove è stata applicata la metodica analitica per la determinazione della durrina con tecnica LC-MS/MS [12]. Da questa viene poi calcolata la quantità di HCN come equivalenti, applicando un fattore di conversione (1 g durrina = 86,7 mg HCN) [13].

INTERVENTI TERAPEUTICI

A. Essendo il caso avvenuto durante il fine settimana, è stato difficoltoso reperire farmaci appropriati. Trenta animali hanno ricevuto in un unico bolo, 4 L per animale, per via endovenosa (e.v.) lenta, differenti soluzioni reidratanti (Ringer lattato,

SintomatologiaEsito esami necroscopici

Decubito laterale, prevalentemente destro (Fig. 1)Forte odore dolciastro proveniente dalle cavità

DispneaSangue di color rosso scarlatto (Fig. 2)

ScialorreaPresenza di schiuma nel lume tracheale

Stupore, eccitazione, fascicolazioniEdema tracheale

Fame d’ariaEnfisema polmonare diffuso

Da lieve a moderato timpanismoAbomasite a sfondo emorragico (Fig. 2)

IpertermiaNecrosi gelatinosa sottocutanea

NistagmoSoffusioni e striature a livello dei prestomaci

MidriasiIdropericardio

RantoliiSoffusioni e petecchie a livello miocardico (Fig. 2)

Splenomegalia con gemizio

Epatomegalia con congestione

Congestione renale

Enterite a sfondo emorragico

Tabella 1 - Elenco dei principali sintomi riscontrati negli animali intossicati e delle osservazioni riscontrate durante gli esami necroscopici.

soluzione fisiologica, glucosata) associate a 60 mL di multivitaminico. Considerate le temperature atmosferiche superiori a 38°C, si è provveduto a rinfrescare gli animali con acqua degli abbeveratoi mobili presenti in campo. Il trattamento è andato a buon fine per 26 capi.

B. Cinque animali sono stati trattati con successo per via e.v., ciascuno con 4 L di soluzione fisiologica contenenti 5 g di tiosolfato di sodio (Na2S2O3). Altri 15 g di Na2S2O3 sono stati disciolti in 10 L d’acqua fredda e fatti assumere per via orale per mezzo di apposite siringhe [14].

C. Considerata l’ora tarda della notte si è tardato a recuperare il Na2S2O3. Ognuno dei 30 animali è stato trattato con 4 L di soluzione reidratante in cui si erano disciolti 30 g di blu di

metilene, iniettata per via e.v.. Una volta giunto in azienda il Na2S2O3, ne sono stati somministrati a ciascun animale 5 g disciolti in sacche di 4 litri di soluzione reidratante per via e.v.. Ventotto capi sono sopravvissuti, 2 sono deceduti la settimana successiva.

D. Non è stato effettuato alcun intervento sugli animali colpiti.

E. Quaranta animali sono stati trattati con successo. Si è effettuato lo stesso trattamento già eseguito nel caso B.

RISULTATI

Esame necroscopico

Gli esami necroscopici hanno evidenziato più o meno uniformemente le lesioni presenti in Tabella 1.

Analisi durrina

Gli esiti degli esami per la determinazione di durrina sono riportati in Tabella 2.

CONCLUSIONI

L’anamnesi, la sintomatologia e la rapida ed elevata mortalità dei bovini intossicati hanno fin da subito suggerito l’avvelenamento da glucosidi cianogenetici. È noto come la terapia consigliata [15] consista innanzitutto nella somministrazione di un agente (nitrito di sodio, in subordine blu di metilene ad alte dosi) in grado di determinare la formazione di metaemoglobina (emoglobina ossidata, incapace di trasportare O2 ai tessuti); tale emoproteina (come la citocromo-ossidasi) è in grado di “catturare” il -CN ripristinando la respirazione cellulare. A questo trattamento deve essere associata la somministrazione di Na2S2O3, che reagisce con il -CN formando tiocianato (-SCN), privo di tossicità acuta. I rischi legati all’impiego di agenti metemoglobinizzanti (la metaemoglobina è incapace di trasportare O2 ai tessuti) e le difficoltà di approvvigionamento legate all’eccezionalità degli episodi e al periodo stagionale hanno di fatto determinato l’impiego quasi esclusivo di tiosolfato sodico. Tale farmacosi è confermato un ottimo antidoto per i bovini nei confronti delle tossicosi da glucosidi cianogenetici, con efficacia del 100% nei due casi in cui è stato tempestivamente somministrato.

Le analisi chimiche hanno rilevato elevata presenza di durrina in 4 casi su 5, risultante in una concentrazione di HCN ben

CASODATALUOGOMATRICEDURRINAHCN

Tabella 2 - Valori di durrina riscontrati nei campioni di sorgo prelevati in ciascuna azienda. I valori di HCN sono stati calcolati come equivalenti a partire dalla concentrazione misurata di durrina.

Figura 1 - Fotografie degli animali intossicati - in decubito laterale destro. Caso A (sopra) e caso B (sotto).

174Outbreaks of lethal cyanogenic glycosides poisonings of cattle after ingestion of Sorghum ssp. grown

oltre la soglia considerata letale di 200 ppm [16]. La Direttiva 2002/32/CE pone a 50 ppm il limite di HCN in materie prime e mangimi [17]. Nel caso E, unico con concentrazioni relativamente basse, è verosimile che il campionamento effettuato sul pascolo di notevole estensione, circa 4 ha, non sia stato sufficientemente rappresentativo.

È interessante notare come la varietà ibrida di sorgo Suzy implicata nel caso A sia commercializzata appositamente per consumo zootecnico. Le indicazioni per l’uso al pascolo consigliano di usare la pianta al di sopra di 80 cm, senza riferimenti specifici a possibili rischi di intossicazione [18]. Per quanto riguarda la sorghetta, l’esame della letteratura ci porta a concludere che si tratti dei primi casi di avvelenamento riportati in Europa.

ARPA Piemonte ha collocato l’estate 2022 tra le più anomale degli ultimi 30 anni [19]. Il bilancio idrico risultava in deficit già dall’inverno precedente, con riserve nivali quasi inesistenti. Le precipitazioni, irregolari sia per quantità che distribuzione territoriale, sono diminuite del 50-60% rispetto agli anni precedenti, specialmente nelle zone colpite dove, inoltre, si è avuto un aumento del numero di notti tropicali (T > 20°C) e giorni tropicali (T > 30°C). Tali condizioni possono dunque aver stimolato un’eccessiva produzione di durrina nel sorgo, causandone un notevole accumulo. Il caso A, ove la cultivar di sorgo era destinata all’alimentazione animale, dimostra quanto sia importante tenere in considerazione le variabili metereologiche per la gestione del sorgo coltivato da foraggio al fine di prevenire il rischio di avvelenamento. In un’ottica di prevenzione del rischio, gli altri casi riportati suggeriscono cautela anche nella gestione del S. halepense, considerata la facile disponibilità di questa specie durante i mesi estivi e il suo utilizzo come foraggio verde storicamente diffuso tra gli allevatori.

RINGRAZIAMENTI

Si ringrazia per la pronta collaborazione in campo i Medici Veterinari Dr. Francesco Barberis dell’ASL di Saluzzo, e il Dott. Francesco Acciardi dell’ASL di Bra. Si ringrazia per la disponibilità data per i campionamenti l’allevatore Giacomino Olivero.

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2.Shapter FM, Crowther A, Fox G, Godwin ID, Watson-Fox L, Hannah IJC, Norton SL (2018) The domestication, spread and uses of sorghum as a crop - Capitolo 2 in: Rooney W (Ed.) “Achieving sustainable cultivation of sorghum - Volume 2: Sorghum utilization around the world”. 1st edition. Burleigh Dodds Science Publishing, London, 31-64.

3.Nahrstedt A (1985) Cyanogenic compounds as protecting agents for organisms. Pl. Syst. Evol. 150, 35-47.

4.Bhat BV (2019) Breeding Forage Sorghum - Chapter 11 in: Aruna C, Visarada KBRS, Bhat BV, Tonapi VA (Eds.) “Breeding Sorghum for Diverse End Uses”. Woodhead Publishing, 175-191.

5.Maffei M (2013) Metaboliti secondari e difese delle piante - Capitolo 9 in: Elementi di Fisiologia Vegetale - Edizione ridotta di: Taiz L, Zeiger E (2010) “Plant Physiology”. 5th edition. Piccin Nuova Libraria S.p.A., Padova, 221-241.

6.Gleadow RM, Ottman MJ, Kimball BA, Wall GW, Pinter PJ, LaMorte RL, Leavitt SW (2016) Drought-induced changes in nitrogen partitioning between cyanide and nitrate in leaves and stems of sorghum grown at elevated CO2 are age dependent. Field Crops Research 185, 97-102.

7.Mandrone M, Chiocchio I, Barbanti L, Tomasi P, Tacchini M, Poli F (2021) Metabolomic Study of Sorghum (Sorghum bicolor) to Interpret Plant Behavior under Variable Field Conditions in View of Smart Agriculture Applications. J. Agric. Food Chem. 69, 1132-1145.

8.Panter KE (2018) Cyanogenic Glycoside-Containing Plants - Chapter 64 in: Gupta RC “Veterinary Toxicology: basic and clinical principles”. 3rd edition. Elsevier -Academic Press, 935-940.

9.Stichler C, Reagor JC (2001) Nitrate and Prussic Acid Poisoning. Texas Agricultural Extension Service, The Texas A&M University System, L-5231 06-01.

10.Nicholson SS (2007) Cyanogenic plants - Capitolo 67 in: Gupta RC (Eds.) “Veterinary toxicology: basic and clinical principles”. 1st edition. Elsevier - Academic Press

11.Varietà ibrida Suzy, Sorghum bicolor (L.) Moench x S. sudanense - registrata in Ungheria, mantenuta da Alfaseed Kft. , Budapest. https://www.niab.com/oecdv2/variety/details/100558/suzy/0/0/vs/ALL/ALL/0

12.Zhong Y, Xu T, Chen Q, Li K, Zhang Z, Song H, Wang M, Wu X, Lu B (2020) Development and validation of eight cyanogenic glucosides via ultra-high-performance liquid chromatography-tandem mass spectrometry in agri-food. Food Chemistry 331, 127305.

13.EFSA Panel on Contaminants in the Food Chain (CONTAM), Schrenk D, Bignami M, Bodin L, Chipman JK, del Mazo J, Grasl-Kraupp B, Hogstrand C, Hoogenboom L (Ron), Leblanc J-C, Nebbia CS, Nielsen E,

Figura 2 - Sangue di colore rosso scarlatto (sinistra), emorragie nell’abomaso (centro) e subepicardiche (destra).

Ntzani E, Petersen A, Sand S, Vleminckx C, Wallace H, Benford D, Brimer L, Mancini FR, Metzler M, Viviani B, Altieri A, Arcella D, Steinkellner H, Schwerdtle T (2019) Evaluation of the health risks related to the presence of cyanogenic glycosides in foods other than raw apricot kernels. EFSA Journal 17(4), e05662.

14.Stöber M (2004) Malattie degli organi respiratori, del diaframma e del torace - Capitolo 5 in: Sali G “Medicina Interna e Chirurgia del Bovino” - Edizione italiana di: Dirkse G, Grunder H-D, Stober M (Ed.) “Innere Medizin und Chirurgie des Rindes” (2002) 4th edition. Le Point Vétérinaire Italie srl, 271-356.

15.Gensa U (2019) Review on Cyanide Poisoning in Ruminants. Journal of Biology, Agriculture and Healthcare Vol.9, No.6.

16.Carlson MP, Anderson B (2013) Cyanide Poisoning. NebGuide - University

of Nebraska-Lincoln Extension, Institute of Agriculture and Natural Resources, G2184.

17.European Parliament, Council of the E.U. (2002) DIRECTIVE 2002/32/EC OF THE EUROPEAN PARLIAMENT AND OF THE COUNCIL of 7 May 2002 on undesirable substances in animal feed. Official Journal of the European Communities, L 140/10.

18.SUZY - Sorgo foraggero multi sfalcio da fasciato - trinciato. Zanandrea Sementi SRL, Bolzano Vicentino (VI). https://zanandreasementi.it/prodotti/suzy-sorgo-foraggero-multi-sfalcio-da-fasciato-trinciato/

19.ARPA Piemonte (2022) Rapporto Siccità in Piemonte 2022 - A cura del Dipartimento Rischi Naturali e Ambientali, Torino. https://www.arpa.piemonte.it/pubblicazioni-2/relazioni-tecniche/analisi-eventi/eventi-2022/rapporto-siccita-in-piemonte-2022

Treatment

of an atresia ani et recti and urogenital agenesis in a calf N

BUSRA KIBAR KURT1, DVM, PHD

SUMMARY

The purpose of this study was to describe the surgical correction of anorectal and urogenital agenesis in a Holstein calf. Clinical examination revealed the absence of anal and urinary openings, testicles or vulvar openings, and suggested the possibility of atresia ani et recti. The complete blood count, blood gas analysis, and biochemistry were evaluated and revealed the presence of a mild respiratory acidosis with a pH of 7.29 and pCO2 level of 62.7 mmHg. The surgery was performed under general anesthesia. During the intraoperative exploration, it was noted that the internal genital organs were not developed and there was a band-like structure between the urinary bladder and the blind pouch colon. Defecation was achieved by performing a ventral colostomy. The band between the urinary bladder and the colon was separated from the colon, a Foley catheter was passed through it by blunt dissection, and it was made to act as the urethra. By using the band-like structure and the Foley catheter, a urethral opening was fashioned and placed caudal to the colostomy opening, which resulted in successful urination. A follow-up checkup after one week showed the calf had no urination or defecation problems and was in good condition. Four weeks later, the owner reported that the calf was doing well and gaining weight. Early surgical treatment can lead to successful outcomes in cases of intestinal atresia and urethral agenesis. The condition known as the imperforate anus, or atresia ani occurs when the anal membrane and perineal skin fail to break down during development, either alone or in combination with other defects. The aim of this study was to present a case of extraordinary atresia ani et recti and urogenital agenesis and its surgical treatment. In small farms, even one calf is important. It should also be considered that the breeder’s emotional attachment to the calves may lead to a preference for treatment.

KEY WORDS

Anorectal anomaly, bovine, colostomy, intestinal atresia, urethral agenesis.

INTRODUCTION

According to the World Health Organization, congenital defects can be defined as abnormalities that occur during intrauterine life. Congenital anomalies can be detected prenatally, at birth, or after birth 1. Intestinal atresia, a congenital defect, has been reported in humans and various domestic animals such as pigs, sheep, and calves. In calves, this abnormality results from abnormal development of the intestinal wall and can affect the rectum/anus, colon, ileum, or jejunum. Intestinal atresia results in a complete blockage of the intestinal lumen, preventing the normal movement of intestinal contents and the passage of fecal material 2,3. Obstruction can occur in different parts of the intestine, such as the duodenum, jejunum, ileum, colon, or anus.

Corresponding Author: Busra Kibar Kurt (busrakibar@yandex.com) (busra.kibar@adu.edu.tr)

ntestinal atresia is a common congenital defect in calves 4,5. Atresia ani et recti is a congenital abnormality with a lack of anal opening and rectum. Vulvar and urethral anomalies are rare. The urorectal septum divides the cloaca into a ventral and a dorsal part. With the differentiation of the cloacal folds, the ventral part forms the urogenital system and the dorsal part forms the digestive system. Urethral agenesis occurs as a result of malformation of this region 6-8. This paper reports an extraordinary case of atresia ani et recti associated with urethral and genital agenesis.

MATERIALS AND METHODS

A 1-day-old Holstein calf (42 kg) was brought to the faculty hospital with a history of not passing meconium or urine since birth. The calf was quite active. Clinical examination confirmed the absence of anal and urinary openings, and testicles or vulvar openings (Figure 1A, B). There was no perineal swelling on

1 University of Aydin Adnan Menderes, Faculty of Veterinary Medicine, Surgery Department, Aydın, Türkiye

178Treatment of an atresia ani et recti and urogenital agenesis in a calf

abdominal compression. The absence of swelling in the anal region strengthened the possibility of it being a case of atresia ani et recti. The body temperature of calf was 38.5°C measured from the mouth. Heart rate was 128/min and respiratory rate was 37/min. The abdomen was slightly distended, but there were no signs of colic. Complete blood count, blood gas, and biochemical analysis and results are detailed in Table 1. Mild respiratory acidosis was observed (pH:7.29, pCO2: 62.7 mmHg).

Meloxicam was administered 0,5 mg/kg (Maxicam, Sanovel, Türkiye). The surgical treatment was performed under general anesthesia. Propofol was administered at 5-6 mg/kg (Propofol-PF 2%, 20ml, Polifarma, Türkiye) by intravenous infusion. Then, endotracheal intubation was performed and 2-3% isoflurane (Isoflurane Usp 100%, 100ml, ABD) was used for inhalation anesthesia. For surgery, the calf was positioned in dorsal recumbency and abdomen was prepared for ventral midline laparotomy. The incision was started caudal to umbilicus to reach vesica urinaria. Intraoperative exploration revealed that the internal genital organs were not developed and there was a band-like structure between the urinary bladder and the end of the colon (Figure 2). The colon was ending as blind pouch. An enterotomy was performed approximately 4-5 cm in front of the blind end of the colon. The band-like structure extending from the vesica urinaria and a 4-5 cm colon section were used for the urethral opening. The enterotomy side was irrigated with a warm saline. A Foley catheter was passed through the colon section to the urinary bladder with blunt dissection and the cuff of the Foley catheter was inflated (Figure 3A, B). The wall of the colon section was sutured to the skin with simple interrupted sutures (USP:0, Monosorb, Sutures) to act as a urethra. The Foley catheter was removed from the abdominal wall caudal to the colostomy opening and urination was achieved. Ventral colostomy was performed. The colon was sutured to the abdominal wall and skin with simple interrupted sutures (USP:0, Monosorb, Sutures) to avoid peritonitis (Figure 4). In the first postoperative week control, it was determined that the calf did not have urination and defecation problems and was in good general condition (Figure 5). When the owner of the patient was contacted by phone 4 weeks later, it was learned that the calf was in good condition and had gained weight.

LaboratoryFindingsReferance

LYM%43.5 %45-75

MON%1.3 %2-7

NEU%55.2 %15-65

RBC x1012/l10.895-10

HGB11.18-15

HCT%40.05 24-46

MCV3740-60

MCHC27.830-36

PLT x109/l289100-800

ALT: Alanine aminotransferase, AST: Aspartate aminotransferase, ALP: Alkaline Phosphatase, GGT: Gamma glutamyl transferase, pH: Actual blood pH, pCO2: Partial pressure of carbon dioxide, pO2, Partial pressure of oxygen, K+: Potassium, Na+: Sodium, Cl-: Clor, WBC: White blood cells; LYM: Lymphocytes, MON: Monocytes, NEU: Neutrophils, RBC: Red blood cells, HGB: Haemoglobin, HCT: Hematocrit, MCV: Mean corpuscular volume, MCHC: Mean corpuscular haemoglobin concentration, PLT: Platelet.

Table 1 - Laboratory data of calf.

Figure 1 - Caudal (Left) and ventral (Right) view of calf, absence of anal and urethral openings.

Figure 2 - Star: Vesica urinaria. Arrowhead: Colon. Arrow: Bandlike structure.

DISCUSSION

Atresia ani and urogenital agenesis develop during the embryonic stage as a result of malformation of the cloacal folds. Atresia ani is a developmental anomaly caused by an autosomal recessive gene. Atresia ani et recti may be associated with recto-vaginal fistula, vagino urethral agenesis, absence of tail,

and diphallus 9-11. In this case, the reason could not be certain. The calf was suffered from atresia ani et recti, uretrhral agenesis and absence of the genital organs. This con-

hypospadias

Figure 5 - Postoperative first week follow up.

Figure 4 - Ventral colostomy (yellow arrow) and urethral opening (black arrow).

Figure 3 - A: Enteretomy side was restricted with penses. B: Foley catheter was reached to vesica urinaria. Star: Vesica urinaria. Yellow arrow: band-like structure. Black arrow: The portion of the colon that remains connected to the band-like structure after enterotomy. Arrowhead: Colon.

180Treatment of an atresia ani et recti and urogenital agenesis in a calf

dition is a rare occurrence 8,12

Previous studies have reported atresia of vagina-urethra and anus 13 in a hybrid calf, atresia ani with recto vestibular fistula and vulvar agenesis in buffalo calf 14, in another buffalo calf atresia ani et recti with agenesis of the vulva and terminal urethra 8, and agenesis of the vulva with atresia ani-et-distal recti in a heifer calf 7. Vahar et al 15 reported a case of colonobladder fistula in a lamb. In the present case, there was a band-like structure between the colon and the urinary bladder, but this structure was not a channel like a fistula. Studies have reported that in cases of intestinal and urethral agenesis, the patient can survive with early surgical treatment. Surgical treatment should be performed before the condition leads to death from autointoxication, circulatory failure, or fecal peritonitis following intestinal and/or vesica urinary rupture 4,7,8,16. When the calf was brought to the clinic at 1 day of age, blood analyses and general condition were found suitable for surgical treatment. Surgical correction was performed under general anesthesia. The patient awoke from the anesthesia without any problems, and in the follow-up 1 week later, the general condition of the patient was good. When the owner of the patient was contacted by phone 1 month later, he reported that he could not bring the calf, but that the calf was healthy.

Due to high frequency of multiple malformations and the role of genetic inheritance in calves with intestinal atresia, it is recommended not to breed these animals, as continued breeding of such animals may have an impact on the persistence of the malformation in the population; even some researchers even recommend the euthanasia for these calves 16. In the present study, the patient was only a livestock animal, as the calf had no genital organs. Patients often do not accept euthanasia, especially because they are more emotional about calves. In this case, the owner of the patient stated that he wanted treatment without considering the cost.

Because atresia ani is often associated with one or more malformations, a careful and complete physical examination should be performed. If perineal bulging does not occur with abdominal pressure, atresia recti should be suspected in addition to atresia ani. Atresia ani et recti with urogenital agenesis are rare cases. Surgical treatment should be aimed at achieving urination and defecation. Treatment should be done in the first few days before problems with food and urea accumulation occur.

Conflict of Interest

The authors declare no conflicts of interest related to this report.

References

1. WHO Congenital anomalies. [Online] World Health Organization.

2. Mee J.F. (2021). Congenital Defects in Calves. In: Szenci O, Mee J, Bleul U, Taverne M, editors. Bovine Prenatal, Perinatal and Neonatal Medicine. 67 Budapest: Hungarian Association for Buiatrics 2021;

3. Keane O.M., Carthy T.R., Hanrahan J.P., Matthews D., McEwan J.C., Rowe S.J., et al. (2023). Risk factors for, and genetic association with, intestinal atresia in dairy calves. Animal Genetics. 54(2), 104-112.

4. Azizi S., Mohammadi R., Mohammadpour I. (2010). Surgical repair and management of congenital intestinal atresia in 68 calves. Veterinary Surgery. 39(1), 115-120.

5. Kaya M., Okumuş Z., Doğan E., Çetin E.M., Yanmaz L.E. (2011). Erzurum Yöresindeki Buzağılarda Doğmasal Anomalilerin Görülme Sıklığı ve Sağkalım Oranları. Fırat University Veterinary Journal of Health Sciences. 25(2), 83-93.

6. Vianna M.L., Tobias K.M. (2005). Atresia Ani in the Dog: A Retrospective Study. Journal of the American Animal Hospital Association. 41(5), 317-322.

7. Wamaitha M.N., Kihurani D.O., Kimeli P., Mwangi W.E., Mande J.D. (2015). Surgical management of agenesis of the vulva with atresia aniet-distal recti in a heifer calf: A case report. Journal of Advanced Veterinary and Animal Research.

8. Hari Krishna N.V.V., Devi Prasad V., Mallikharjuna Rao C. (2009). Agenesis of vulva and terminal urethra with atresia ani et recti in a buffalo calf. Buffalo Bulletin. 28, 165-167.

9. Loynachan A.T., Jackson C.B., Harrison L.R. (2006). Complete diphallia, imperforate ani (type 2 atresia ani), and an accessory scrotum in a 5day-old calf. Journal of Veterinary Diagnostic Investigation. 18(4), 408412.

10. Bademkiran S., çen H., Kurt D. (2009). Congenital Recto Vaginal Fistula with Atresia Ani in a Heifer : A Case Report. Y.Y.U. Vetriner Fakulties Dergisi. 20, 61-64.

11. Köm M., Eröksüz Y. (2013). Urorectal septum malformation sequence in a calf. Kafkas Universitesi Veteriner Fakultesi Dergisi. 19(SUPPL.A).

12. Tyagi R., Singh J. (1999). Ruminant Surgery. 222, New Delhi, India: CBS Publishers;

13. Shetty B., Nigam J., Singh G. (1978). Surgical correction of vagino-urethral and anal agenesis in a calf. Indian Veterinary Journal. 55, 234-236.

14. Sreenu M., Rao M., Naidu K., Rao T. (1998). Atresia ani associated with rectovestibular fistula and vulvar agenesis in a buffalo calf. Buffalo Bulletin. 17(2), 41-42.

15. Vahar M., Hosseni S.M., Omidzahir S., Kenari E.O., Iraee M.A., Ziabari A. reza H. (2015). Report of congenital colonobladder fistula with atresia ani in a lamb and treatment by surgery. Asian Pacific Journal of Tropical Disease. 5(S1), S181-S183 Asian Pacific Tropical Medicine Press;

16. Meylan M. (2008). Surgery of the Bovine Large Intestine. Veterinary Clinics of North America - Food Animal Practice. 24(3), 479-496 Elsevier Ltd;

ÖMER TARıK ORHUN1, YAKUP KOCAMAN*1, SıTKıCAN OKUR1, LATIF EMRAH YANMAZ2, CIHAN ÖZ3,

1 Department of Surgery, Faculty of Veterinary Medicine, Atatürk University, Erzurum, Turkey

2 Department of Surgery, Faculty of Veterinary Medicine, Burdur Mehmet Akif Ersoy University, Burdur, Turkey

3 Department of Microbiology, Faculty of Veterinary Medicine, Atatürk University, Erzurum, Turkey

SUMMARY

Septic arthritis, defined as joint inflammation, is common in newborn calves when the passive transfer of maternal immunoglobulins fails. An unhygienic environment, lack of umbilical care and insufficient colostrum after birth predispose to the formation of septic arthritis. Arthrotomy and through-and-through needle joint lavage techniques are employed in the management of septic arthritis. This study aimed to evaluate the effect of two different joint lavage techniques on survival rate in calves with septic arthritis presented to Atatürk University Veterinary Faculty Animal Hospital between 2018-2021. A total of 248 calves with septic arthritis were included in the study. One hundred twenty (48.4%) of the 248 calves had a complete bacterial analysis of the infected joint. Two different lavage methods (through-and-through needle [n = 138] and arthrotomies [n= 96]) were used for joint lavage. A through-and-through lavage method was performed on 138 calves. Out of these, 114 (82.6%) animals made a full recovery, while 24 (14.4%) were humanely euthanized or died. Conversely, arthrotomy was employed in 96 calves, with 81 (84.4%) animals making a full recovery, while 15 (15.6%) calves were humanely euthanized or died. Additionally, Of the 248 calves, 120 (48.5%) obtained a thorough bacterial examination of the affected joint. Eighty-nine out of 120 (74.1%) calves had at least one bacterial isolate found during bacterial culture. Two bacterial isolates were discovered from synovial samples of 31 (25.8%) calves. Staphylococcus aureus (30%), Escherichia coli (25.8%), Trueperella pyogenes (24.1%), and other pathogens (13.4%), including Enterococcus species, Bacillus species, Proteus species, and Corynebacterium species, were the most commonly isolated bacteria in the infected joint of calves. There was no statistically significant difference in the survival rates between the two joint lavage techniques (P = 0.72). The chances of success of treatment are related to whether the patient has been treated before, and the stage of the disease, but both treatment methods can be used successfully in the treatment of arthritis since the success rates are more or less the same.

KEY WORDS

Calves, Septic arthritis, Joint, Hematogenous.

INTRODUCTION

Septic arthritis is a very important disease that frequently affects newborn calves, especially within the first eight weeks of their life. It is thought to be the result of secondary bacterial colonization, especially in the joints affected by the disease. Especially common in newborn calves and is defined as a joint disease that causes by direct trauma, hematogenous dissemination, and extension of periarticular infections. It is a common cause of lameness, which is responsible for significant economic losses in the livestock and dairy industries (1,2). Systemic infections in calves may also be a cause of septic arthritis (2,3). Also, it can result from periarticular wound infection, inoculation of the joint by an open or puncture wound, and by

Corresponding Author: Yakup Kocaman (yakup.kocaman@atauni.edu.tr)

hematogenous or lymphomatous inoculation. The higher incidence of septic arthritis in calves is due born in an unhygienic environment, lack of disinfection of the umbilicus, and inadequate intake of colostrum in the first few hours after birth. An inflammatory response occurs in the infected joint. Microorganisms are destroyed by neutrophils and their enzymes such as collagenase, cathepsin, elastase, and gelatinase that come to the site (4,5). Which enzymes cause damage to joint components and cartilage tissues (6).

Early treatment in calves with septic arthritis significantly affects the prognosis of the disease. As the disease progresses, the chances of success decrease. Several methods for septic arthritis treatment have been described. Antibiotic and anti-inflammatory therapy, joint lavage, arthroscopy, arthrotomy, and arthrodesis methods are used in the treatment of septic arthritis. If there is no recovery in the joint after medical therapy, surgical methods may be a good option. Joint lavage is an effective invasive method, the application of which is based on surgical asepsis procedures in cases that do not respond to par-

MÜMIN GÖKHAN ŞENOCAK1, MUHAMMET FURKAN KAPLAN3, UĞUR ERSÖZ1, ECE AYTEK3

Comparison of the effect of two different joint lavage techniques on survival rate in calves with septic arthritis: 248 cases N

182Comparison of the effect of two different joint lavage techniques on survival rate in calves with septic arthritis: 248 cases

enteral antibiotic therapy (2,7). Arthrotomy, like arthroscopy, is indicated in very serious and chronic cases where joint lavage fails or medical treatment has a low chance of success. Surgical opening of the joint may be required when the presence of fibrin formations blocks. Surgical management serves as a supplement to medical treatment and attempts to relieve synovial pressure and bacterial strain (8). Joint lavage is aimed at irrigation and evacuation of infected contents in cases of purulent-septic arthritis. In chronic cases, joint lavage is insufficient, as the joint is often filled with compact fibrin. The prognosis is poor in cases with soft tissue, bone necrosis, and osteomyelitis. In such cases, the recommended treatment options can be listed as arthroscopy, arthrotomy, or arthrodesis (6,9,10,11).

Clinical signs of septic arthritis include lameness, distension, increasing heat, redness, and signs of pain during palpation of the affected joints. The diagnosis of septic arthritis is confirmed based on arthrocentesis findings (bacterial culture results, cytological assessment of synovial fluid, and biomarker measurement), ultrasonographic and radiographic examination of the affected joints (1,12).

There are insufficient data on the comparison of the arthrotomy and through-and-through joint lavage methods of septic arthritis in calves. This study aimed to compare the effect of arthrotomy and through-and-through joint lavage techniques on survival rate in 248 newborn calves with septic arthritis.

MATERIALS AND METHODS

The medical record database of the Atatürk University Veterinary Faculty of Animal Hospital was reviewed to identify the records of calves that were examined from 2018 through 2021 for which septic arthritis was diagnosed at ≤ 90 days old. Cases were divided into treatment groups according to clinical, radiographic, and ultrasonographic examination findings. The

exclusion criteria are specified in Table 1.

Clinical Examination

The general condition of the presented animals is slightly and moderately impaired. After the inspection and palpation examination, treatment options were applied as indicated in Table 1.

Radiographic Examination

A radiographic examination was performed on the anteriorposterior and latero-medial positions of the infected joints. For the examination were used a stationary X-ray machine (Mex100, Oberhausen-Germany). In acute cases, the presence of radiolucent area in the joint and increased joint space were observed. In chronic cases, decreased joint gap, bone lysis on the subchondral joint surface, periosteal reaction, and bone proliferation were observed.

Ultrasonographic Examination

The affected joints were shaved, cleaned, and contact gel was used before the ultrasonographic examination. Joints were investigated by using a 5 MHz sector and 7.5 MHz linear probes (GE Vivid S6, Milwaukee, USA). No anesthetic drug was used during both examinations. In some cases, on ultrasonographic examination, the joint contents were seen as hyperechogenic and anechogenic areas. Again, in cases, fibrin structures were determined from mild to severe.

Microbiological Examination

Before the infected joint treatment, calves were administered with intramuscular 40 µg/kg detomidine (Domosedan, Zoetis, NJ, USA). The affected joint was prepared for aseptic conditions using standard sterilization procedures. A sterile puncture of the affected joint using a 16-gauge needle was performed and 2 ml synovial fluid was collected with a 2.5 ml syringe. To determine the presence of Mycoplasma spp. in the samples,

- Clinical radiographic and ultrasonographic findings of 248 newborn calves with septic arthritis.

they were inoculated into Mycoplasma selective medium. The medium contains yeast extract, thallium acetate, penicillin G, and phenol red solutions. For routine isolation of Mycoplasma, the specimen should be inoculated into broth and agar. The inoculated medium (broth and agar) is incubated in a humid atmosphere at 37 C, under 5% CO2, and 48 and 96 hours. The Petri dishes are viewed under a stereoscopic microscope or the low-power objective of a light microscope, for the characteristic mycoplasmal ‘fried-egg’ microcolonies. The cultures can be regarded as negative if no microcolonies are seen after 14 days of incubation.

For routine diagnosis, two milliliters of synovial fluid were inoculated onto Columbia agar with 7% defibrinated sheep blood and MacConkey agar. Specimens plates were incubated at 35 ± 2 C in a normal atmosphere for 24 to 48 hours. The morphological and hemolytic characteristics of the colonies, catalase, coagulase, and oxidase tests, gram staining, and specific biochemical assays were used to identify any growth on agars. According to the Clinical Laboratory Standards Guide, antibiotic susceptibility testing was performed using the Agar Disc Diffusion technique. The colonies were suspended in a 0.9% NaCl saline solution. Turbidity was corrected to the 0.5 McFarland standard (approximately 108 CFU/mL) and utilized as the antibiotic test inoculum. Following that, 0.1 mL of bacterial solution was put over Muller Hinton agar, followed by antibiotic disks. Disks containing Amoxicillin-clavulanic acid (20/10 g), ampicillin-sulbactam (10/10 g), streptomycin (10 g), tetracycline (30 g), trimethoprim (5 g) were collected from Oxoid® (Hampshire, England) and plates were incubated at 37 °C for 24 h. According to the Clinical Laboratory Standards Guide, inhibition zones were assessed and classified as susceptible, moderately resistant, or resistant.

Treatment

Medical treatment

Amoxicillin-clavulanic acid (10 mg/kg, Intramuscular (IM), q 24 h) or Marbofloxacin (2 mg/kg, IM, q 24 h) were administered for antibiotic therapy for 5 days until culture results were obtained. For anti-inflammatory effects, Flunixin meglumine (2.2 mg/kg, IM, q 24 h) was applied for 3 days during the treatment.

Through-and-Through Joint Lavage

The a-16-gauge needle was inserted into the two opposite points of the involved joint until reaching the synovial membrane. Then, a large volume of NaCl 0.9% (Bioflex, Istanbul, Turkey) solution was used for joint lavage. To create pressure in the joint, the synovial sac was inflated at some times by grabbing the caudal outlet. The joint contents were completely evacuated. In cases deemed necessary, the procedure was repeated 2 to 3 times. Joint lavage was continued until the aspirated fluid becomes clear. No bandage was applied after the procedure.

Arthrotomy

The animals were placed on the operating table in the lateral position. The involved joint was shaved circularly. The joint area was washed to remove solid dirt. Then, povidone-iodine was used for antisepsis. Detomidine (Domosedan, Zoetis, NJ, USA) at a dose of 80 mcg/kg was used for sedation, and intravenous (IV) 2 mg/kg ketamine hydrochloride (Ketasol, Interhas, Turkey) was used for general anesthesia. General anesthesia was maintained with boluses of ketamine hydrochloride

IV administered every 20 minutes at the initial dosage. The procedure was started with a 1-2 cm incision using the number 12 scalpel of the relevant joints. After skin and muscle incisions in the region, the synovial membrane was incised to approach the joint. Fibrinous or purulent contents and destroyed bone fragments in the infected joint were removed by gentle curettage. The joint chambers were debrided continuously with isotonic saline solution throughout the entire procedure. After arthrotomy, the incision sites were not closed to allow drainage. The area was protected with a window bandage. During this procedure, the tendons, adjacent ligaments, vessels, and nerves were protected from trauma.

Outcome

The calves were not hospitalized. If the patient performed following the owner’s expectations 1 year after discharge from the hospital, the long-term result was considered positive. This information was obtained by a researcher by calling the patient owners by phone. The result was defined as negative if the calf died in the hospital or afterward, was euthanized, or did not meet the owner’s expectations during the telephone survey.

Statistical Analysis

All data were analyzed using Medcalc version 20.011 (Medcalc Software, Ostend, Belgium) and values of <0.05 were considered significant. The results of different lavage techniques on survival rate were also compared by chi-square analysis. The results of the treatment of calves with septic arthritis are presented.

RESULTS

The study was conducted on 248 cases. All of the cases were evaluated clinically, radiographically, and ultrasonographically as in Table 1. Joint radiographs were determined abnormal in 182 (73.4%) cases, including periarticular soft tissue swelling, decrease or increase in joint space, bone proliferation, and subchondral lysis. The ultrasonographic examination was performed in 40 (16.1%) calves and inflammatory effusions with varying echogenicity-echogenicity were found.

A total of 234 animals were treated with either thorough-andthrough lavage or arthrotomies. A through-and-through lavage method was performed on 138 calves. Out of these, 114 (82.6%) animals made a full recovery, while 24 (14.4%) were humanely euthanized or died. Conversely, arthrotomy was employed in 96 calves, with 81 (84.4%) animals making a full recovery, while 15 (15.6%) calves were humanely euthanized or died. There was no statistically significant difference in the survival rates between the two joint lavage techniques (P = 0.72). Twelve animals (4.8% of the total 248) were excluded from the study groups due to severe calcified joint content, advanced bone destruction, and refusal of treatment by the animal owners. One hundred twenty (48.4%) of the 248 calves had a complete bacterial analysis of the infected joint. During bacterial culture, eighty-nine of 120 (74.1%) calves had at least one bacterial isolate identified. Thirty-one (25.8%) calves had two bacterial isolates identified from synovial specimens. The most isolated bacteria in the infected joint of calves was Staphylococcus aureus (30%), followed by Escherichia coli (25.8%), Trueperella pyogenes (24.1%), and other pathogens (13.4%) such as Enterococcus spp., Bacillus spp., Proteus spp., and Corynebacterium spp

184Comparison of the effect of two different joint lavage techniques on survival rate in calves with septic arthritis: 248 cases

DISCUSSION

Septic arthritis, commonly observed in calves, results from umbilical disease or deficiency of passive transfer of maternal immunoglobulins after birth or postnatal trauma (1). The current study evaluated that calves aged less than 3 months with septic arthritis presented to Ataturk University Animal Hospital were described. Lameness, swelling, pain, hyperthermia, and loss of appetite are common clinical findings in calves with septic arthritis (1,6,13). In this study, all of these clinical findings were present.

A total of 248 calves were diagnosed using ultrasonography or radiography examination in this study. Although radiography and ultrasonography are considered specific in the diagnosis of septic arthritis, no abnormalities were detected on ultrasonography or radiography of the joints of 20 calves with septic arthritis. The most common radiographic findings of infected joints were increased or decreased joint space, bone proliferation, periarticular soft tissue swelling, and subchondral lysis in this study, and were similar to those reported in the previous studies in calves with septic arthritis (7,14,15).

Joint lavage is an important treatment method in the treatment of septic arthritis. Aggressive antibiotic therapy, along with joint drainage and/or lavage can be effective in early cases, but arthrotomy is considered to give better results in any chronic cases or severe infections (16). In this study, the through-and-through joint lavage method was used in calves aged 1-2 weeks and the arthrotomy method was used in calves aged 2-12 weeks. Experimental studies have shown that fibrin clots develop in the joint after the seventh day (17). But depending on the severity of the disease, it may be necessary to use different lavage methods. The most effective way to remove the formed fibrin clots is arthrotomy. Arthrotomy allows precise debridement of tissues inside the joint, and removal of infected synovia, bone, and bone fragments. The success of the arthrotomy method is higher than other applications. Therefore, in this study, the arthrotomy method was preferred in animals aged 2-12 weeks with dense fibrin accumulation. In severe cases, total curettage and arthrodesis performed to ensure immobilization of the joint give successful results. But joint immobilization is not recommended in young animals (17). Arthroscopy, which is another surgical method, is limited in chronic cases. Because the fibrin in the joint can impair vision and manipulation. On the contrary, arthrotomy provides better access to all joint faces. Increases the success in the removal of exudate and necrotic tissues. However, the surgical approach is difficult because of its proximity to adjacent tissues such as muscles, vessels, and nerves (2).

As reported in previous studies (13,18), at least one bacterial pathogen was identified in joints with septic arthritis in 120 of the 248 (48.4%) calves in the current study. The most isolated bacteria in infected joints were Staphylococcus aureus (30%), followed by Escherichia coli (25.8%), Trueperella pyogenes (24.1%), and other pathogens (20.1%) in calves, which seems similar to bacteriological findings in previous studies (13,19).

The treatment of septic arthritis includes reducing the bacterial load and preventing the increase in inflammation, and pain management. The bacterial load, the duration of the affected joint, and the disease are important for determining case management (1). The successful treatment for septic arthritis is the removal of purulent debris from the infected joint. There are

several techniques described in calves including tidal irrigation, through-and-through needle lavage, performing arthrotomies, or flushing the joint under arthroscopic guidance. As mentioned in previous studies, needles were inserted into the bulging joint while applying joint through-and-through joint lavage. A previous study conducted on humans with septic knee arthritis reported that needle aspiration and arthrotomies method did not cause a change in survival rate (20). Similarly, the current study stated that there was no difference between through-andthrough needle lavage and arthrotomies in the survival rate. A lot of biomarkers have been studied in vacationing the diagnosis and prognosis in cases of septic arthritis (1). Although bacterial culture and antibiogram analysis were performed in our study, synovial fluid analysis, hematological analysis, and total nucleated cell count analyses from synovial fluid could not be performed.

In this study, the survival rate of through-and-through joint lavage and arthrotomy, which are one of the treatment options, were evaluated on calves. No significant difference was found between the effect of the two methods on survival rate. As a result, the success rate of joint lavage methods in the treatment of calves with septic arthritis is increased by early diagnosis and treatment. The choice of treatment method should be decided by clinical, radiographic, and ultrasonographic examination findings. The chance of success of the treatment is related to the stage of the disease, whether the patient has been treated before or not, and scientific studies should be conducted on more animal numbers and species to compare the success of these two different treatment methods.

Author’s contribution

ÖTO, YK, and SO conceptualization, investigation, data curation, writing-original draft preparation, formal analysis; LEY, MGS and UE, supervision, writing – review & editing, methodology, investigation, visualization; CÖ, MFK, and AE supervision, visualization, and microbiological analysis.

Funding

This research received no external funding.

Declaration of Competing Interest

The authors declare no conflict of interest.

Acknowledgment

The authors thank Ayse Golgeli and Ferda Turgut for their advice and support in conducting the research.

References

1.Jackson P. (1999). Treatment of septic arthritis in calves. In Practice, 21: 596-601.

2.Desrochers A., Francoz D. (2014). Clinical management of septic arthritis in cattle. Vet. Clin. N. Am. - Food Anim. Pract., 30:177-203.

3.Heppelman, M., Kofler J., Meyer H., Rehage J., Starke A. (2009). Advances in the surgical treatment of septic arthritis of the distal interphalangeal joint in cattle. J. Vet. Med., 182: 162-175.

4.Kofler J., Martinek B. (2005). New surgical approach to the plantar fetlock joint through the digital flexor tendon sheath wall and suspensory ligament apparatus in cases of concurrent septic synovitis in two cattle. J. Vet. Med., 169, 370-375.

5.Weaver AD. (1997). Joint conditions. In: Lameness in Cattle, Ed. Greenough, P.R. and Weaver, A.D, 3rd. ed., 277-293. W.B. Saunders Co, Philadelphia.

6.Köstlin R.G., Nuss K. (1988). Treatment of septic pedal arthritis in cat-

tle by joint resection results, Tierärztl Prax, 16: 123-131.

7.Desrochers A. (2004). Septic Arthritis. In: Farm Animal Surgery, Ed. Fathman E.Z., 2nd ed., 330-336, Elsevier Sci., Philadelphia.

8.Jackson P. (1999). Treatment of septic arthritis in calves. In Pract., 21: 596601.

9.Mulon P.Y., Desrochers A., Francoz D. (2016). Surgical management of septic arthritis. Vet. Clin. N. Am. - Food Anim. Pract., 32: 777-795.

10.Verschooten F., Vermeiren D., Devriese L. (2000). Bone infection in the bovine appendicular skeleton: a clinical, radiographic, and experimental study. Vet Radiol Ultrasound, 41: 250-260.

11.Steiner A., Hirsbrunner G., Miserez R., Tschudi P. (1999). Arthroscopic lavage and implantation of gentamicin-impregnated collagen sponges for treatment of chronic septic arthritis in cattle. Vet Comp Orthop Traumatol., 12: 64-69.

12.Constant C., Masseau I., Babkine M., Nichols S., Francoz D., Fecteau G., Desrochers A. (2018). Radiographic study of hematogenous septic arthritis in dairy calves. Vet Comp Orthop Traumatol., 31: 252-260.

13.Starke, A., Kehler, W., & Rehage, J. (2006). Arthrotomy and arthrodesis in the treatment of complicated arthritis of the fetlock joint in adult cat-

tle. Vet. Rec., 159: 772-777.

14.Vasanthkumar H.G., Narayanan M.K., Sudheesh S., Sreeranjini A.R., Devanand C.B. (2018). Evaluation and Management of Septic Arthritis in Calves: a review of six cases. J. Vet. Anim. Sci, 49: 70-73.

15.Dogan E., Yanmaz L.E., Okumus Z., Kaya M., Gokhan M., Cengiz S. (2016). Radiographic, ultrasonographic, and thermographic findings in neonatal calves with septic arthritis: 82 cases. Ataturk Univ. Vet. Bilim, 11: 6-12.

16.Munroe G. A., Cauvin E. R. (1994). The use of arthroscopy in the treatment of septic arthritis in two Highland calves. Br Vet J., 150: 439-449.

17.Duncan, J. R., Mahaffey, E. A., Prasse, K. W. (1994). Veterinary laboratory medicine (Vol. 243).,3rd ed., 37-62, Iowa State University Press, Ames

18-.Bailey J. V. (1985). Bovine arthritides: Classification, diagnosis, prognosis, and treatment. Clin. North Am. Food Anim., 1: 39-52.

19.Goodarzi M., Khamesipour F., Mahallati S.A., Dehkordi M.K., Azizi S. (2015). Study on prevalence of bacterial causes in calves arthritis. J. agric. biol. Sci., 10: 206-212.

20.Pawasuttikul C. (2013). Comparison of needle aspiration and arthrotomy treatment for septic knee arthritis: A 10-year retrospective study. JseaOrtho., 37: 29-33.

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Systemic embolic hepatitis and pneumonia caused by subacute ruminal acidosis in

Korean indigenous cattle: A case report

1 Department of Veterinary Internal Medicine, College of Veterinary Medicine, Jeonbuk National University, Iksan 54896, Republic of Korea

2 Department of Animal Hospital, Hanwoo (Korean indigenous cattle) genetic improvement center, National Agricultural Cooperative Federation, Seosan 31948, Republic of Korea

3 Department of Animal Science and Biotechnology, College of Ecology and Environmental Science, Kyungpook National University, Sangju 37224, Republic of Korea

SUMMARY

A 44-month-old Korean indigenous cow was found in the lateral recumbency in a pasture while grazing. Decreased heart sound and rate, hypothermia, scleral injection, subcutaneous petechial hemorrhage, and moderate dehydration were observed upon inspection. Complete blood count showed significant neutropenia. Serum biochemical analysis revealed azotemia, increased globulin, decreased albumin/globulin ratio, and increased aspartate transaminase, creatinine kinase, phosphorus, and magnesium. Blood gas analysis indicated hypokalemia and high anion gap metabolic acidosis. Ultrasonography of the right abdomen and chest showed hyperechoic nodules in the liver and cranial lung fields.Based onclinical status, late state sepsis was diagnosed, and necropsy was performed after euthanasia. Embolic inflammation was found in the liver, lung, and spleen, rumenitis and ruminal parakeratosis were observed, and the pH of the recovered ruminal fluid was 5.5. Based on necropsy findings, death for sepsis, systemic embolic hepatitis and pneumonia caused by subacute ruminal acidosis (SARA) was assessed. While SARA is thought to be prevalent in Korean indigenous cattle raised with concentrated feed, few studies or case reports that would confirm this have been published to date.

KEY WORDS

Embolic inflammation; Korean indigenous cow; subacute ruminal acidosis ultrasonography.

INTRODUCTION

Ruminal acidosis can be clinically divided into acute and subacute acidosis (SARA)1. Both forms can be attributed to the excessive supply of concentrated feed and low-quality roughage14. While acute forms can be caused by sudden oversupply, subacute forms differ inasmuch as they arise from chronic oversupply1-4. Therefore, the acute type is considered an individual disease, but the subacute type can be considered a herd disease5

Acute ruminal acidosis has to be considered an emergency. Moderate dehydration is observed and, unless properly treated, mortality rate can reach 90%3. With SARA, in contrast, there are no obvious clinical signs of dehydration or overt symptoms, but the disease affects other organs, such as liver, also causing a decrease in productivity2-4.

A prerequisite for acute and subacute ruminal acidosis is a de-

Corresponding

Author:

Jinho Park (jpark@jbnu.ac.kr)

crease of ruminal pH. During acute rumen acidosis, excessive lactic acid production by microflora increases osmotic pressure and fluid loss, resulting in systemic severe dehydration and systemic lactic acidosis1. The primary cause of SARA is an increase in volatile fatty acids (VFAs) that decrease ruminal pH6 Under an acidic environment, ruminal epithelium unprotected by the mucus develops secondary ruminitis7. The acidic environment in the rumen causes the death of Gram-negative bacteria, release of Lipopolysaccharide (LPS) and leads to a systemic inflammatory response8, 9. Liver abscesses, pneumonia, endocarditis, pyelonephritis, and arthritis can also be caused by the migration of bacteria into the rumen through blood vessels, and caudal vena cava thrombosis can also occur2-5 Based on the results of previous studies in which a sensor was inserted a sensor in the rumen, Korean indigenous cattle seem not to be resistant to SARA10. Since Korean indigenous cattle are raised on concentrated feed, SARA may be prevalent, and this is potentially major factor for decreased productivity of livestock farms in Korea. To the best of our knowledge, this is the first report of SARA with embolic hepatitis and pneumonia detected by ultrasonography in Korean indigenous cattle.

YOUNGJUN KIM1,2, JI-YEONG KU1, KYOUNG-SEONG CHOI3, AND JINHO PARK1*

188Systemic embolic hepatitis and pneumonia caused by subacute ruminal acidosis in Korean indigenous cattle

CASE PRESENTATION

A 44-month-old Korean indigenous cow was observed in lateral recumbency in a pasture while grazing. Physical examination showed hypothermia (38°), a respiratory rate of 20 beats/min, and a lowered heart rate of 37 beats/min with dampened heart sounds. Moreover, scleral injection and petechial hemorrhage on the pinna were found. Based on this information, the animal was clinically diagnosed to be in later stages of sepsis. After hematological analysis and ultrasonography, this animal was necropsied because of extremely severe condition. A complete blood cell (CBC) test using a Procyte DX hematologic analyzer (IDEXX Laboratories, Westbrook, ME, USA) revealed increased hematocrit (Hct, 49.5%) and a decrease in white blood cells count, especially neutropenia (0.02 x 103/µL). Blood gas analysis (i-STAT 1 analyzer, Abbott, USA) showed high anion gap metabolic acidosis. Serum biochemistry analysis (Drychem 4000i, Fujifilm, Japan) revealed a decrease in the albumin/globulin ratio (0.53) and an increase in globulin (5.1 g/dL), suggesting chronic inflammation. Moreover, creatinine was 2.7 mg/dL, and blood urea nitrogen (BUN) was 103.4 mg/dL, indicating azotemia. Aspartate aminotransferase (AST) was also increased to 328 U/L, and indirect bilirubin was 1.2 mg/dL with a total bilirubin of 1.3 mg/dL, indicating hepatic-predominant hyper-bilirubinemia. Lactate level was very high at 11.08 mmol/L (Table 1).

An ultrasound examination (LOGIQ e R8, GE Healthcare, USA) using a 5 MHz linear probe revealed many hyperechoic nod-

Figure 1 - Ultrasonogram of right abdomen and chest. (a) hyperechoic nodules were found in the parenchyma of the liver at the right 9th and 10th intercostal spaces. (b) At the right third intercostal space, consolidation and a hyperechoic nodule were found in the lung parenchyma. The depth of the consolidation in lung was 4.73 cm and the mean diameter of the nodules was 0.46 cm.

ules in the right 9th, 10th, and 11th intercostal space (Figure 1a). In the right third intercostal space, consolidation was found in the right cranial ventral lobe of the lung and multiple hyperechoic nodules with a mean diameter of 0.46 cm (Figure 1b). The echogenicity of the pancreas observed in the right 13th intercostal space was significantly more hyperechoic than nor-

Table 1 - Findings on serum chemistry and blood gas analysis in a Korean indigenous cow with systemic embolic inflammation caused by SARA.

but no liver abscess was found. (c

normal.

mal. However, the diameter of the caudal vena cava was normal.

Consequently, decreased heart rate and sound on auscultation, hypothermia, scleral injection, and marked neutropenia on the CBC test, and high anion gap metabolic acidosis accompanied by a rapid increase in lactate on blood gas analysis were observed. This was assumed to be the end stage of septic shock, and euthanasia followed by necropsy was performed for the animal’s welfare.

Necropsy revealed marked jugular vein dilation, and embolism with splenomegaly (Figure 2a). In consistent with ultrasound findings, embolic hepatitis without abscess was observed (Figure 2b). Although lungs were diffusely bright red, some congestion was found in the cranial lobe of the lung, indicating embolic pneumonia (Figures 2c and 2d). Moreover, erosion and parakeratosis due to inflammation were observed in the rumen wall (Figure 3), and the pH of the ruminal fluid collected through the necropsy was acidified to 5.5. Therefore, embolic systemic inflammation in the liver, lungs, and spleen, as well as sepsis, was caused by chronic SARA.

DISCUSSION

Studies in grazing cattle in Ireland and New Zealand11, 12 have shown that SARA is caused by a diet with high fermentable and

Figure 2 - Gross findings after euthanasia. (a) Embolic inflammation with splenomegaly. (b) Embolic hepatitis in the posterior lobe of the liver,

and d) Although embolic pneumonia was found throughout the lung field, the overall lung color was

Figure 3 - A number of erosive findings (arrow) in rumen were found due to ruminal acidosis.

190Systemic embolic hepatitis and pneumonia caused by subacute ruminal acidosis in Korean indigenous cattle

low fiber grass12, 13. In our case, beef cattle used for breeding in four month of grazing and in the fourth month of pregnancy, 6 months after parturition, were examined. It is not clear what caused SARA; some factors might have been the increase in their feed intake in the one month immediately post-calving, the intake of grasses that were lush and low fiber in improperly managed pastures, or some combination of these factors.

A diagnosis of SARA is usually confirmed by a ruminal pH of 5.5 or less14, 15. In this case, the pH was 5.5. The average pH of cattle fed mainly concentrated diet is 5.5-6.5, while the average pH of cattle fed with forage is 6.0-7.23, 16. Thus, while a pH of 5.5 corresponded to the baseline, the result was considered particularly meaningful considering that forage represented the main feed in grazing pasture. In addition, one report mentioned that the risk of SARA exists when the ruminal pH is 5.5-5.817 Perhaps a new diagnostic criterion for ruminal pH in cattle reared with concentrated feed and roughage should be established. In addition to the evaluation of ruminal fluid by oral probe or rumenocentesis, the evaluation of SARA by measuring rumen wall thickness using transabdominal ultrasonography is have been studied, which is likely to be utilized in the future18. Unlike the abomasum, ruminal mucosa protective factors do not exist; so it can be chemically damaged by acidic ruminal fluid7. These factors directly cause rumenitis and reduce the integrity of rumen epithelial cells19. Gram-negative bacteria sensitive to low pH die, releasing LPS, which are introduced (along with ruminal bacteria) into systemic circulation through the portal vein. These can then cause inflammation in organs such as liver, lungs, and kidneys2-5, 8, 9. However, in this case, no liver abscess, which is commonly found in cases of SARA, and no caudal vena cava syndrome, were observed. Moreover, rumenitis and ruminal parakeratosis were observed in this case which is caused by hyperproliferation of the ruminal epithelial cells due to increased VFAs such as butyric acid and propionic acid20. Ruminal parakeratosis has also been reported to further accelerate rumenitis by interfering with the absorption of VFAs21.

Ultrasonography to evaluate liver disease in cattle has been previously reported, specifically for liver abscess and fatty liver2224. In the case of liver abscesses, nodules can vary from nonechoic to hyperechoic, and from homogeneously to heterogeneously25. Ultrasound evaluation of pulmonary lesions, such as the presence of pulmonary abscess through measurement of diameter of mass or nodule, and the presence of pulmonary consolidation through identification of Broncho-aerogram and fluid bronchogram with evaluation of echogenicity of lung parenchyma, have been reported26, 27. Several hyperechoic nodules, similar to those found in the liver were also found in the lungs (the right 9th, 10th, and 11th intercostal spaces). These findings suggest that embolic inflammation should be considered as a differential diagnosis when there are multiple nodules in various organs on ultrasonography. In cases of septic shock, heart rate and heart sound increase in the early stage (Hyper-dynamic phase), whereas heart rate and heart sound decrease is seen in the late stage when blood flow to tissues drops off (Hypo-dynamic phase)3. Our case showed a decreased respiratory rate (20 beats/min) and heart rate (37 beats/min) with dampened heart sound. Multiple scleral injections and petechial hemorrhages were also observed. These findings indicated the late stages of septic shock. We note, however, that a definition of septic shock applicable

to cattle has not yet been established. Fecteau G et al. defined the condition with any two criteria of the following: blood culture growth within 48 h, bacteria cultured as pathogens, white blood cell increase/decrease in CBC, fibrinogen of 500 mg/dL, and more than 2% of band neutrophils were observed28. As sepsis is an emergency, considering the significant time it would take to perform all these tests, we suggest that methods for rapid diagnosis of sepsis in cattle should be developed. In this case, the lactate level had increased to 11.08 mmol/L. As tissue hypoxia caused by sepsis can increase lactate levels3, this may prove valuable as a diagnostic tool for sepsis.

Acknowledgements

Not applicable

Funding

This study was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, and Forestry (IPET) (grant No. 122017-02-1-HD020). This research was partially supported by a National Research Foundation of Korea (NRF) grant funded by the Korean government (MSIT) (No. 2021R1A2C100517111).

Conflict of Interest

The authors declare that there were no conflicts of interest.

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28. Fecteau G., Pare J., Van Metre D.C., Smith B.P., Holmberg C.A., Guterbock W., et al. (1997). Use of a clinical sepsis score for predicting bacteremia in neonatal dairy calves on a calf rearing farm. Can Vet J, 38:101-104.

Pre-maxilla

Ó

JENA BISWADEEP1, ANAND ARUN2 AND SANGWAN VANDANA3*

1 PhD Scholar, Department of Veterinary Surgery and Radiology, College of Veterinary Science, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, 141004, Punjab, India.

2 Professor, Department of Veterinary Surgery and Radiology, College of Veterinary Science, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, 141004, Punjab, India.

3 Associate Professor, Department of Veterinary Surgery and Radiology, College of Veterinary Science, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, 141004, Punjab, India.

SUMMARY

The present report describes the successful surgical management of rostral pre-maxilla fracture using inter-fragmental stainless steel wire in a 6 month old colt foal. The foal had the history of bite on the premaxilla by another horse on the day of presentation. The clinical examination and lateral skull radiograph confirmed open, complete fracture at rostral premaxilla (incisive bone) region with displacement. The fracture fixation was done under general anaesthesia in dorsal recumbency. The fracture was primarily stabilized with a two 2 mm Steinmann pins followed by tightening an inter-fragmentary 20-gauge stainless steel wire in a horizontal mattress formation with the ‘U’ part on hard palate and free ends on the outside of incisors. One month follow up revealed radiological and clinical fracture union and the wire was removed under general anaesthesia.

KEY WORDS

Foal, Horse bite, Pre-maxilla, Rostral, Stainless Steel wire.

INTRODUCTION

Fractures of rostral premaxilla are common in foals and are mostly due to kick by another animal or, self-inflicted trauma1 and are usually presented at the diastema region vis-à-vis transverse2-4. More complicated fractures of the face involving sinuses and multiple bones or comminuted fractures have been reported in equine5. Internal fixation is mostly recommended for such fractures to reinstate normal occlusion and stability that can facilitate fracture healing. Wiring is the most suitable repair technique2-4 described for pre-maxilla fractures in foals but plating alone or plating as an external fixator6 has been described for comminuted fractures of mandible in adult equines. The present report describes the successful surgical management of open, rostral pre-maxilla fracture using inter-fragmental stainless steel wire in a foal with a history of horse bite on the day of presentation.

CASE HISTORY AND PRESENTATION

A 6-month-old colt foal, weighing 150 Kg, was presented with a history of bite on the premaxilla by another horse in early

Corresponding Author: Vandana Sangwan (drvandanasangwan@rediffmail.com).

morning hours. The clinical examination revealed an open (Figure 1) fracture at rostral premaxilla (incisive bone) region. All the other physiological parameters were normal. Tetanus toxoid was injected to the foal.

RADIOGRAPHY

The lateral skull radiograph confirmed a simple, complete fracture at rostral premaxilla (incisive bone) region with displacement (Figure 2). The foal was decided for surgical intervention under general anaesthesia with the consent of the owner.

SURGICAL TREATMENT

The foal was pre-anaesthetized with injection Xylazine hydrochloride @ 1.1mg/Kg, IV followed by injection Ketamine hydrochloride @ 2.2mg/Kg, IV at 4 minutes. The foal lied down and was intubated with ID 18 endotracheal tube. The foal was hoisted to operation table and was placed in dorsal recumbency. The general anaesthesia was maintained with Isoflurane mixed in 100% oxygen. Intravenous fluids (Inj. Normal saline @ 10ml/Kg) were started and first antibiotic injection of Ceftriaxoneand Tazobactam, 1.5g was given IV.

The fracture site was prepared antiseptically using 1% betadine

fracture repair using inter-fragmentary stainless steel wire in a foal

in normal saline. There was no loss of any incisor of premaxilla. The fracture was primarily stabilized by drilling two, 2 mm Steinmann pin in the inter-alveolar space of teeth 101-102 and 201-202 (Figure 3a). One, 2 mm Steinmann pin was removed and a hypodermic 16-gauge needle was inserted into the drilled hole. This 16-gauge needle was used as a cannula to facilitate passage of 20-gauge stainless steel wire. The same procedure was repeated on the contralateral drilled pin site inside out. This helped in formation of a horizontal mattress pattern with the ‘U’ part of the wire on the hard palate (caudal fracture fragment) and the free ends on the outside of the incisors. The free ends of the wires were twisted together to reduce and compress the fractured fragments (Figure 3b). After tightening, the ends of the wires were cut and flattened against the gum (Figure 3c). Intraoral lacerated soft tissue lesion at fracture site was closed using #2 polyamide sutures (Sutupak®, Ethicon) (Figure 3d). The immediate post-operative radiograph showed satisfactory apposition (Figure 3e).

POST OPERATIVE CARE AND FOLLOW UP

Post-operatively, the foal was advised antibiotics injection Ceftriaxone+ Tazobactam 1.5 g twice daily for 05 days, IV and analgesic injection Flunixin meglumine 3ml, IV for 3 days. A soft palatable diet (green grass) was advised for 15 days. The 28 day follows up revealed the clinical (Figure 4a) and radiological (Figure 4b) fracture union. The wire and the remaining PDS sutures were removed under general anaesthesia (Figure 4c). The foal underwent uneventful recovery.

DISCUSSION

Premaxilla fractures in foals are common due to the kick and not bite by another horse. The premaxilla fractures, if presented early and stabilized adequately are reported to heal fast with good outcome due to abundant vascularization of gingiva1-4, 7. Multiple fracture fixation methods have been reported to repair premaxilla including the tension band wiring, pinning, plates, screws, splintage (extraoral or, intraoral), but majority were amenable to wiring alone. Wiring was considered comparatively simple but provided satisfactory stability with no requirement for specialized or expensive equipment2-4 General anaesthesia is required while repairing fractures in equine and xylazine sedation followed by ketamine induction was found satisfactory for this foal. However, different combinations for foals and adult equines have been described in literature6

The cranial fractures of mandible and maxilla can be evaluated without radiography8 but if available, radiography may help in diagnosing any other fracture line in the vicinity. The choice of higher generation antibiotic was made in the present case to avoid any complications of root tooth infection, secondary sinus infection pertaining to wire or osteomyelitis4,9,10 which otherwise might lead to significant morbidity. Equine being a grazing animal and the upper lip being the major prehensile organ, the premaxilla is exposed to continuous shearing force during eating/ grazing. Therefore, biomechanical properties of fracture fixation methods bear paramount importance. The complications and even re-fracture of premaxilla have been reported in 68% of equine1-3, 7. But the present technique was sufficient to provide biomechanical stability and thus, resumption of normal feed intake without any complications.

CONCLUSION

Inter-fragmentary stainless wiring is a simple, efficient and in-

expensive technique to provide adequate rostral pre-maxilla fracture stability using minimal equipment in foal. The fragments may be pre-stabilized with intramedullary pins which can be removed after placing the wire.

Conflicts Of Interest

The authors have no conflicts of interest with anyone.

Authors Contribution

Author 1 wrote the manuscript and was assisting surgery. Author 2 was the major surgeon; Author 3 was the radiologist and anaesthesiologist.

Acknowledgement

The authors are thankful to Rashtriya Krishi Vikas Yojana (RKVY) and Guru Angad Dev Veterinary and Animal Sciences University (GADVASU), Ludhiana, Punjab, India for their support and cooperation in carrying out the study.

References

1.Henninger R. W., Beard W. L., Schneider R. K., Bramlage L. R., Burkhardt, H. A. (1999). Fractures of the rostral portion of the mandible and maxilla in horses: 89 cases (1979-1997). J Am Vet Med Assoc, 214(11): 16481652.

2.Wilson D. A. (2011). How to Repair Rostral Mandibular and Maxillary Fractures. Proceedings Am Assoc Equine Pract, 57: 290-295.

3.Çetinkaya M. A., and Demirutku A. (2012). Interfragmental fixation of rostral mandibular fracture with cerclage wire in a thoroughbred English horse. Turk J Vet Anim Sci, doi: 10.3906/vet-1102-792

4.Naddaf H., Sabiza S., Kavosi, N. (2015). Surgical treatment and a unique management of rostral mandibular fracture with cerclage wire in a horse. Vet Res Forum, 6(2): 181-183.

5.Gardner A. K., Santschi E. M., Mudge M. C., Belknap J. K., Metzler A. G. (2020). Intrasinus bolstering of traumatic maxillary sinus fractures by using Foley catheter balloons in two foals. Vet Surg, 49(6):1255-1261. doi: 10.1111/vsu.13433

6.Mählmann K., Cender A. N., Ehrle A., Lischer C. J. (2021). Application of locking compression plates as type 1 external fixators to treat unilateral mandibular fractures in four equids and one dromedary. Vet Surg, 51 (2): 341-352. doi.org/10.1111/vsu.13748

7.Ramzan P. H. L. (2008). Management of rostral mandibular fractures in the young horse. Equine Vet Edu, 20(2): 107-112. doi: 10.2746/095777308X272094

8.Schumacher J. Treatment of horses for injuries of the head. Western Veterinary Conference; Las Vegas, USA. 2013. p. EQ41.

9.Boorman S., Lascola K., Mora-Pereira M., Ceriotti S., Boone L. (2020). Internal Fixation of a Complicated Mandibular Fracture in a Filly Using a String-of-Pearls Locking Plate Assisted by a 3D Printed Model. Vet Comp Ortho Traumatol (open), 3 (2): e90-95. doi: 10.1055/s-0040-1716351

10.Çetinkaya M, Demirutku A. Interfragmental fixation of rostral mandibular fracture with cerclage wire in a thoroughbred English horse. Turk J Vet Anim Sci. 2012; 36 (1):67-71.

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