Antibodies to parainfluenza virus type 3 in goat population in Poland.

Respiratory diseases constitute a major health problem in small ruminant herds around the world, and parainfluenza virus type 3 (PIV-3) has been shown to play a vital role in their etiology. This cross-sectional study describes the serological status of the non-vaccinated dairy goat popu- lation in Poland with respect to PIV-3 infection and investigates the relationship between the presence of antibodies to PIV-3 and some basic herd-level and animal-level factors, including small ruminant lentivirus (SRLV) infection. Serum samples from 1188 goats from 48 herds were tested for the concentration of antibodies to PIV-3 using a quantitative immunoenzymatic assay. Specific antibodies were detected in all tested goats from all herds. The concentration of PIV-3 antibodies varied from 8.4 to >240 ng/ml (median 95.9 ng/ml) and was significantly higher in goats from larger herds and from these herds in which cough was often observed by farmers. Moreover, it was noted that female goats had higher antibody concentrations than males. On the other hand, the concentration of PIV-3 antibodies did not prove to be significantly linked to the presence of SRLV infection. This study shows that PIV-3 infection in the Polish goat population is widespread and appears to contribute to the occurrence of respiratory diseases in goat herds.

Nasal carriage of various staphylococcal species in small ruminant lentivirus-infected asymptomatic goats.

The study was carried out in Polish goat population to estimate the prevalence of the nasal cavity infection with various staphylococcal species including methicillin-resistant Staphylococcus aureus(MRSA), investigate the potential permissive role of small ruminant lentivirus (SRLV) infection and determine the level of clonality of S. aureus nasal isolates. Nasal swabs and blood samples were collec-ted from 1300 clinically healthy adult goats from 21 Polish goat herds. Blood samples were serological-ly screened for SRLV. Staphylococci were isolated from nasal swabs and identified using classical microbiological methods, MALDI-TOF, multiplex-PCR, and their clonality was assessed using PFGE. Antimicrobial resistance was determined on the basis of minimum inhibitory concentration and by demonstration of the presence of the mecA gene encoding the multiplex-PCR PBP2a protein and of the five main types of staphylococcal cassette chromosome mec. The apparent prevalence of staphylococ-cal and S. aureus infection of the nasal cavity was 29.1% (CI 95%: 26.9%, 31.5%) and 7.3% (CI 95%: 6.1%, 8.8%), respectively. No relationship was found between the SRLV-infection and the presence of any staphylococcal species including S. aureus (p=0.143). Only 9.8% of S. aureus isolates were resistant to amoxicillin/clavulanic acid and 5.9% to chloramphenicol and ciprofloxacin. All tested isolates proved to be phenotypically and genotypically sensitive to methicillin, which yielded the appar-ent prevalence of MRSA of 0% (CI 95%: 0%, 7.0%). S. aureus isolates show high genetic similarity within goat herds, however vary considerably between herds. Goats do not appear to be an important source of S. aureus for humans in Poland.

Herd-level seroprevalence of pestivirus infection in goat population in Poland.

A disease survey was conducted between 2014 and 2018 in the Polish goat population to determine the seroprevalence of pestiviral infection. Blood samples from 910 goats (782 females and 128 males) were collected in 62 goat herds and tested for bovine viral diarrhea virus (BVDV) infection with a competitive ELISA in a serial fashion. Between 10 and 13 adult female goats were randomly selected from each herd, assuming individual-level seroprevalence of pestiviral infection in a herd of at least 30% and a level of confidence of 95%. In each herd, all males were tested. At least one seropositive goat was found in each of the 4 herds. However, in one herd, the only positive goat tested negative in serial retesting. Finally, 3 herds were consi-dered as seropositive which yielded apparent herd-level seroprevalence of 4.8% (CI 95%: 1.7%, 13.3%). After adjusting for the ELISA herd-level sensitivity and specificity, the true herd-level prevalence was 3.9% (CI 95%: 1.2%, 12.0%). No males tested positive. In 2 out of 3 seropositive herds, goats regularly shared pastures with cattle.

Development of resistance to eprinomectin in gastrointestinal nematodes in a goat herd with pre-existing resistance to benzimidazoles.

We described a first case of resistance to eprinomectin in goat herd in Poland in which resistance to benzimidazoles had been previously reported. The herd was established in 2011 by purchasing several goats from a single herd in south-eastern Poland. Resistance to benzimidazoles in the herd was first reported in 2017. Shortly after the owner started to signal low effectiveness of the treatment with eprinomectin. In June 2018 the larval development test from pooled faecal sample was performed and the results indicated the presence of resistance to macrocyclic lactones and levamisole. In July 2018 a faecal egg count (FEC) reduction test was performed in 39 animals with levamisole, eprinomectin and one untreated control group. Drugs were used in doses recommended for goats. Three methods of calculation of FEC reduction were compared. After eprinomectin treatment, FEC reduction ranged from 0 to 20%, depending on the method of calculation. FEC reduction following levamisole treatment was 100%. Main species present in the faecal samples after treatment and in larvicidal concentrations in larval development test was Haemonchus contortus. This is the first report of anthelminthic resistance to macrocylic lactones (eprinomectin) in goats in Poland.

Malignant thymoma - the most common neoplasm in goats.

In the literature the occurrence of thymomas in goats varies from 0.7 to 25%, depending on the study. Therefore the current investigation was carried out to determine the prevalence of thymoma in goats in Poland. Between 2007 and 2018 at the Warsaw Veterinary Faculty 399 goat autopsies and ultrasound examinations of the chest in other 145 goats were performed. Mediastinal tumors were diagnosed during post mortem examination in 2 goats. Additionally, ultrasound examination of the chest revealed a large mass close to the heart in the thoracic cavity in 1 case. This goat was euthanized and an autopsy confirmed a mediastinal tumor. Histopathological examination, with immunohistochemical tests to anti cytokeratin, p63 and p40 confirmed thymomas in all three cases. In our study thymomas were found in 0.5% (95% CI: 0.1% to 1.8%) of examined goats and they represented the most common malignancy in this species.

The concentration of copper, zinc, manganese and selenium in the serum and liver of goats with caprine arthritis-encephalitis.

Concentrations of four trace elements, copper (Cu), zinc (Zn), manganese (Mn) and seleni- um (Se), have thus far proven to be affected by lentiviral infections in people and rhesus monkeys. As small ruminant lentivirus (SRLV) infection is responsible for one of the most important goat diseases, caprine arthritis-encephalitis (CAE), we evaluated serum and liver concentrations of Cu, Zn, Mn, Se in goats severely affected by symptomatic CAE and compared them with litera- ture reference intervals. Serum and liver samples of dairy goats euthanized due to severe clinical form of CAE were collected and screened for the concentration of Cu, Zn, Mn (54 serum sam- ples, 22 liver samples), and Se (36 serum samples, 22 liver samples) using flame atomic absorption spectrometry for Cu, Zn, Mn and graphite furnace atomic absorption spectroscopy for Se. In both serum and liver samples concentration of Zn was the highest, followed by Cu concentration, and then by Mn and Se. There was no relationship between serum and liver concentrations of trace elements. Liver concentrations of all four trace elements and serum Cu concentration fell within literature reference intervals, although liver Se concentration was mainly in the lower marginal range (between 0.4 and 1.0 mg/L). Serum Zn concentration was elevated (>1.2 mg/L) in all goats, serum Mn concentration was elevated (>0.04 mg/L) in 42 (78%) goats and serum Se concentra- tion was elevated (>1.6 mg/L) in 13 (36%) goats. Concluding, severe symptomatic CAE does not appear to be associated with the level of any of the four trace elements.

Diagnostic performance of ID screen MVV-CAEV Indirect Screening ELISA in identifying small ruminant lentiviruses-infected goats.

Diagnostic performance of ID Screen MVV-CAEV Indirect Screening ELISA in identifying goats infected with small ruminant lentiviruses (SRLV) was evaluated. In total 299 serum samples from the collection of the Laboratory of Veterinary Epidemiology and Economics--109 truly positive and 190 truly negative--were used. To be enrolled in the study a serum sample had to come from at least 2 year-old goat which had reacted identically in two serological surveys preceding sample collection and was kept in a herd of stable serological status confirmed at least twice during preceding 5 years. Moreover, in seropositive herds at least 20% of goats had to be serologically positive at the moment when the serum sample was collected for the study. The test proved to have high accuracy. Area under curve was 98.8% (95% CI: 97.5%, 100%). Diagnostic performance of the test was almost identical (Youlden's index of 90%, sensitivity > 90% and specificity > 95%) within a fairly wide range of cut-off values--between 20% and 60%. At manufacturer's cut-off of 50% sensitivity and specificity were 91.7% (95% CI: 85.0%, 95.6%) and 98.9% (95% CI: 96.2%, 99.7%), respectively. For this cut-off positive likelihood ratio was 87 (95% CI: 22, 346) and negative likelihood ratio was 0.08 (95% CI: 0.04, 0.16). In conclusion, the results of this study indicate that ID Screen MVV-CAEV Indirect Screening ELISA is a highly accurate diagnostic test for SRLV infection.

Schmallenberg virus antibodies detected in Poland.

Between 24 and 30 July 2012 230 adult goats from three western provinces of Poland bordering on Germany (Western Pomerania, Lubuskie and Lower Silesia) were blood-sampled and tested for antibodies to Schmallenberg virus (SBV) using indirect immunoenzymatic test (ID Screen® Schmallenberg virus indirect, IDvet Innovative Diagnostics). The ELISA test identified 21 seropositive goats - 15 in Western Pomerania (16% of all goats tested in this province), five in Lubuskie (6%) and one in Lower Silesia (2%). Our study demonstrates for the first time the presence of antibodies to SBV in Poland.

PFGE and AFLP genotyping of Staphylococcus aureus subsp. anaerobius isolated from goats with Morel's disease.

Staphylococcus aureus subsp. anaerobius is the etiological agent of the Morel's disease in sheep and goats. The disease presents with subcutaneous abscesses, located mainly in the superficial lymph nodes. Forty-one isolates of S. aureus subsp. anaerobius were collected from two outbreaks of the Morel's disease in Poland in years 2006-2008. Analysis of DNA SmaI digests by PFGE showed that 35 of 41 isolates belonged to the same PFGE type, identical to the type strain of S. aureus subsp. anaerobius ATCC 35844, confirming high level of clonality of the species. The DNA patterns of the remaining identical 6 isolates, different from the reference strain only by two bands, were found closely related. Genotyping performed with AFLP technique revealed two clonal groups including 16 and 25 isolates, respectively. The study indicated that AFLP technique might be a better discriminatory tool for genetic analysis of S. aureus subsp. anaerobius isolates, when compared to PFGE.

A note on the organization and expression of ß-defensin genes in Polish goats.

Nucleotide (cds) and amino acids sequences of the caprine ß2-defensin genes were in silico compared to search for the sequence variation and for the LAP gene sequences in the goat genome and for the presence of LAP gene transcripts in goat tissues. The comparison of the exon sequences revealed that the first 64 amino acids are identical in both LAP and ß1-defensin. However, the GBD-1 prepropeptide is shorter by 18 amino acids due to the presence of the stop codon UAA at position 209-211 in GBD-1 mRNA. The LAP gene, which was found, so far, only in Indian goat breeds, is absent in the genome of Polish dairy goats. The introns of the caprine ß1- and ß2-defensin genes were, for the first time, sequenced; their sequences showed 99.6 % identity, differing in six nucleotide positions.

Multivariate model for the assessment of risk of fetal loss in goat herds.

The observational study was carried out in a population of Polish breeding goats in 2007 to determine the prevalence of fetal loss and identify risk factors contributing to its occurrence. The multivariate model allowing to predict the risk of the occurrence of fetal loss in a herd in a study population was developed. Data on the occurrence of fetal loss, as well as of 28 hypothesized risk factors were collected from goat owners using standardized questionnaire during face-to-face reviews on farms. Moreover, data on the herd-level seroprevalence of four abortifacient infections--Chlamydophila abortus, Leptospira spp., BVDV-1 and Neospora caninum--were included in the final analysis. Fetal loss was reported as occurring often in 12 of 49 goat herds (24.5%). The relationship between the hypothesized risk factors and the occurrence of fetal loss was verified in the multivariate logistic regression (alpha=0.05). Final analysis yielded four risk factors: regular veterinary supervision at least twice a year (OR 0.188; CI 95% 0.054 - 0.656), frequent occurrence of injuries and fractures (OR 3.172; CI 95% 1.081 - 9.310), frequent occurrence of respiratory signs in adult goats (OR 4.848; CI 95% 1.353 - 17.377) and presence of antibodies to C. abortus in a herd (OR 58.116; CI 95% 1.369 - 2466.438). The accuracy of the multivariate model was analyzed using receiver operating characteristic (ROC) curve technique. Area under the curve was 0.895 (CI 95% 0.801-0.981). For optimal cut-off value of 0.20-0.35 the multivariate model had sensitivity of 75.00% and specificity of 89.19% in predicting fetal loss in a herd.

Effect of isosporiasis prevention with toltrazuril on long-term pig performance.

The efficacy of toltrazuril treatment was assessed in two experiments in Polish swine herds. Experiment 1 included a toltrazuril treatment group, Group A (n = 410), and untreated control, Group B (n = 386). Time to sale in Group A was 108 days versus 120 days for Group B, with average body weights at sale of 114.2 kg and 108.8 kg, respectively (P < 0.05). In experiment 2, the health status and body weight gain of 238 piglets treated with toltrazuril (Group D) were compared to 235 untreated piglets (Group K). A similar difference was observed in average body weights of slaughtered animals, being on average 104 kg in Group D and 101 kg in Group K (P < 0.01). Animals from Group D were slaughtered 5 days earlier than animals from Group K (day 166 versus day 171). Data from clinical trials suggest treatment of coccidiosis with toltrazuril offering potential for improved animal welfare and yields, however this has remained unproven in field conditions in large swine production facilities. The present study confirms the efficacy of toltrazuril treatment when used in the field and the subsequent positive impact on time to weaning, time to market, and on weight gain at all time points.

Twelve-year cohort study on the influence of caprine arthritis-encephalitis virus infection on milk yield and composition.

This long-term observational cohort study was carried out to evaluate the effect of caprine arthritis-encephalitis virus (CAEV) infection on the quantitative and qualitative characteristics of milk production in dairy goats. For this purpose, a dairy herd comprising both CAEV-infected and uninfected female goats was observed for 12 consecutive years. Records on daily milk yield, somatic cell count (SCC), and contents of the major milk components (fat, protein and lactose) were collected every month. In total, 3,042 records (1,114 from CAEV-positive and 1,928 from CAEV-negative animals) from 177 female goats were used for statistical analysis. The multi-trait repeatability test-day animal model using the derivative-free multivariate analysis package with the average information-REML method was applied to eliminate the influence of factors other than CAEV infection on milk production in goats. The statistical significance of the differences between estimates for seropositive and seronegative goats was evaluated using Student's t-test. The effect of age of goats (parity) on their serological status was also estimated with the one-trait repeatability test-day model. The serological status of goats was linked to parity: the higher the parity, the greater the probability of CAEV infection. No significant differences between infected and uninfected goats with respect to daily milk yield and SCC were found. On the other hand, the milk of uninfected goats contained more total protein (3.40% vs. 3.35%), fat (3.69% vs. 3.54%), and lactose (4.30% vs. 4.25%) than the milk of infected goats. Even though these differences were highly significant, they were small when expressed numerically.

Leptospiral antibodies in the breeding goat population of Poland.

In 2007, a survey was conducted on the prevalence of antibodies to 19 Leptospira serovars in goats in Poland . Sera were collected from adult females of all 49 breeding goat herds in Poland by applying simple random sampling. In total, 736 sera were tested by the microscopic agglutination test. A herd was considered seropositive if at least one goat with a titre of 100 or more was detected. Herd-level seroprevalence of Leptospira was 89.8 per cent and individual-level seroprevalence varied from less than 1.0 to 85.0 per cent among the herds. Antibodies to Leptospira serovars Zanoni, Bratislava, Autumnalis, Australis and Javanica were most frequently detected. Although 40.3 per cent of seropositive goats had high antibody titres (≥ 400), suggesting recent infection, no relationship with abortions or other clinical manifestation of leptospirosis in goats was detected.

Evaluation of the risk factors influencing the spread of caseous lymphadenitis in goat herds.

Epidemiological studies on caseous lymphadenitis were carried out in Poland in 1996 and 2002 among goat herds covered by a milk recording program. Between-herd seroprevalence was 13.2% in 1996 and increased to 62.5% in 2002. The average size of seropositive herds was statistically significantly higher than that of seronegative ones, however there was no statistically significant difference in the age between the herds. A statistically significant prevalence ratio (PR) was identified and relevant attributable risk for exposed animals (AR(exp)) was calculated for the following risk factors: presence of seropositive males in a herd (PR = 8.350; AR(exp) = 0.651), presence of superficial abscesses in animals (PR = 6.142; AR(exp) = 0.620), presence of respiratory signs (PR = 2.900; AR(exp) = 0.393), presence of animals in poor condition in a herd (PR = 2.774; AR(exp) = 0.390) and occurrence of reproductive failures in a herd (PR = 1.798; AR(exp) = 0.230). Purchase of animals from abroad, mastitis and husbandry conditions (housing system, grazing system, hygienic conditions) were not shown to be statistically significant risk factors.

Influence of chronic caprine arthritis-encephalitis virus infection on the population of peripheral blood leukocytes.

The influence of caprine arthritis-encephalitis (CAE) virus infection on the population of peripheral blood leukocytes in goats was evaluated. For this purpose two groups of adult dairy female goats were formed. The experimental group consisted of 17 goats, which had been naturally infected for many years. The control group comprised 29 non-infected goats, which originated from CAE-free herd. All goats were clinically healthy. Whole blood was collected and tested in hematological analyzer and light microscope to assess the total number of leukocytes and the percentage of four leukocyte populations--neutrophils, eosinophils, monocytes and lymphocytes. Then, flow cytometry with monoclonal antibodies against several surface antigens (namely CD14, CD2, B-B2, CD4, CD8h, TCR-N6, WC1-N2 and WC1-N3) was performed to assess the proportion of lymphocyte subpopulations. Statistically significant differences (alpha < or = 0.01) were observed only in the subpopulations of T lymphocytes--percentage of all subpopulations were significantly higher in the group of seropositive goats. No statistically significant differences were revealed with respect to the total number of blood leukocytes, the average percentage of blood leukocyte populations and proportions of both T and B lymphocytes.

Prevalence of antibodies against Chlamydophila abortus and Coxiella burnetii in goat herds in Poland.

An epidemiological study was carried out to determine the herd prevalence of Chlamydophila abortus and Coxiella burnetii antibodies in goats covered by a milk recording program in Poland. The survey took place in 2007 and 48 herds located in different parts of the country were involved. A representative sample from each herd was taken by a simple random sampling allowing to detect seropositivity of a herd on a 95% level of confidence. In total 918 goats were tested for specific antibodies against both germs with the use of enzyme-linked immunosorbent assays. In addition, history of reproductive failures was recorded in these herds. The survey revealed that the herd prevalence of C. abortus was 4.2% (2 herds) while no C. burnetii antibodies were found. Abortions were reported to be a problem in 80% of herds while repeating estrus was encountered in 46% of herds. Reproductive failure concerned two seropositive herds as well. Since the germ is present in the population, it has to be taken into consideration in diagnostic process. Nevertheless, the results of the present study indicate that C. abortus infection occurs infrequently in Polish goats. As no antibodies against C. burnetii were detected in the screened sample the risk of goat-to-human transmission of both bacteria in Poland seems to be very low.

Epidemiological features of Morel's disease in goats.

Morel's disease caused by Staphylococcus aureus subsp. anaerobius was diagnosed for the first time in Poland in October 2006 in a goat flock. A second infected flock was found two months later. The course of the disease in both flocks was observed for 15-17 months. Clinical manifestation was confined to abscesses located near major superficial lymph nodes, mostly: superficial cervical, subiliac, parotid and mandibular. At necropsy no other lesions were found. The incubation period was estimated at 3 weeks. Clinical signs were seen both in young and adult goats and up to 7 abscesses in one animal were noted. Abscesses tended to persist for 1 to 5 months, then rupture and heal completely. The initial high in-flock point prevalence in both flocks (93.6% and 84.4%) dropped to approximately 10-30% during next 3-4 months. Until the end of the observation period the in-flock point prevalence remained at this level and only single abscesses were observed, mainly in young animals. No influence of the concurrent caprine arthritis encephalitis virus (CAEV) infection on the clinical course of Morel's disease was noticed. It is to be concluded that the clinical course of Morel's disease in a goat flock resembles caseous lymphadenitis (CLA). However, in Morel's disease abscesses occur more frequently in young goats and are located near, not inside, the lymph nodes, as in the case with CLA. Also, the incubation period of Morel's disease seems to be shorter (3 weeks versus 2-6 months in CLA).

Influence of caprine arthritis-encephalitis virus infection on the activity of peripheral blood lymphocytes.

Non-specific lymphocyte transformation assay using phytohemagglutinin (PHA) as a mitogen was applied to evaluate influence of caprine arthritis-encephalitis virus (CAEV) infection on activity of peripheral blood lymphocytes. Animals were selected for the CAEV-infected and CAEV-non-infected groups according to the results of two serological surveys carried out at one year interval, with the use of an ELISA test. In goats which were not infected with CAEV, lymphocytes stimulation index (SI) revealed a high diversity of the results with an mean value equal to 5.86 (minimum = 0.45, maximum = 40.00, SD = 8.40). SI values for infected goats reached the average of 1.10 (minimum = 0.46, maximum = 1.85, SD = 0.26). The difference between the average lymphocyte stimulation indices was statistically highly significant in both groups (p = 0.002) which could be an evidence of CAEV infection influence on lymphocyte reactivity. Regarding ELISA test as a "golden standard" the application of lymphocyte transformation assay in diagnosis of CAEV infection was assessed. The ROC curve was drawn. The area under the curve was only 0.324, which indicates very low accuracy of this method and limits its use for the diagnosis of the disease.

Serological evidence of lack of contact with caprine herpesvirus type 1 and bluetongue virus in goat population in Poland.

Investigation into herd-level seroprevalence of caprine herpesvirus type 1 (CpHV-1) and bluetongue virus (BTV) was conducted in 2007 in Poland. It involved the entire population of goats covered by a milk recording program in 2007, which included 49 goat herds. The number of goats examined in each herd was determined statistically in order to detect the presence of at least one seropositive animal in a herd with a 95% probability and simple random method of sampling was applied. No antibodies to CpHV-1 or BTV were detected. Further calculations were carried out to determine the herd-level true seroprevalence, taking into account sensitivity and specificity of the test as well as several other factors. It can be concluded that till the middle of 2007 population of Polish goats covered by the milk recording program remained negative with respect to CpHV-1 and BTV.