Lactate dehydrogenase isoenzymes in the lungs of sheep with acute and chronic pneumonia

Total lactate dehydrogenase and the absolute and percentage levels of its isoenzymes were measured in lung lesions and macroscopically normal areas of lung from lambs with chronic proliferative exudative pneumonia and acute pasteurella pneumonia. Lung lesions had a higher total enzyme activity which was associated mainly with increases in the activity of the LDH4 and LDH5 isoenzymes, particularly in chronic pneumonia, and gave lung lesions a considerable potential for altering the serum isoenzyme distribution. Thus, the nature of any changes in the serum isoenzyme distribution will depend on whether the isoenzymes are released from abnormal or normal areas of lung. This appears to be the first report on lactate dehydrogenase isoenzymes in ovine pneumonia.     

Lactate dehydrogenase isoenzymes in the lungs of sheep with acute and chronic pneumonia

Total lactate dehydrogenase and the absolute and percentage levels of its isoenzymes were measured in lung lesions and macroscopically normal areas of lung from lambs with chronic proliferative exudative pneumonia and acute pasteurella pneumonia. Lung lesions had a higher total enzyme activity which was associated mainly with increases in the activity of the LDH₄ and LDH₅ isoenzymes, particularly in chronic pneumonia, and gave lung lesions a considerable potential for altering the serum isoenzyme distribution. Thus, the nature of any changes in the serum isoenzyme distribution will depend on whether the isoenzymes are released from abnormal or normal areas of lung. This appears to be the first report on lactate dehydrogenase isoenzymes in ovine pneumonia.     

Comparison of direct culture versus PCR for the detection of Brucella in aborted fetuses of cattle and sheep in Turkey

The aim of this study was to detect Brucella in samples from aborted fetuses of sheep and cattle in Turkey using PCR and bacteriological analysis, and to determine the sensitivity and specificity of the PCR. Organ homogenates from 38 aborted fetuses of cattle and 56 aborted fetuses of sheep were tested. All organ homogenates were cultured for bacteriological analysis, and all of the homogenates and the Brucella isolates obtained by culture were examined with a commercial PCR kit. On bacteriological analysis, Brucella species were found in 30 (31.9 per cent) of the 94 organ homogenates, eight (21.1 per cent) of which were from cattle and 22 (39.3 per cent) from sheep. Using PCR, a total of 29 (30.9 per cent) homogenates were positive for Brucella species, eight (21.1 per cent) of which were from cattle and 21 (37.5 per cent) from sheep. Compared with the bacteriological method, the diagnostic sensitivity and specificity of the PCR kit used in this study were 83 per cent and 94 per cent, respectively.     

Identification of Corynebacterium pseudotuberculosis isolates from sheep and goats by PCR

The present study was carried out to estimate the prevalence of caseous lymphadenitis (CL) in sheep and goats slaughtered at the local abattoir in Elazig province located in the east of Turkey, between September and December 2000. A total of 2046 sheep and 2262 goat carcasses were examined during the study period and 118 abscessed lymph nodes, 89 from sheep and 29 from goats, were collected. Corynebacterium spp. strains were isolated from 81.4% of the abscesses, giving an overall prevalence of 2.2%. The prevalence was 3.5 and 1.1% in sheep and goats, respectively. PCR on DNA extracted from 96 suspicious isolates, using a pair of Corynebacterium pseudotuberculosis-specific primers, was positive for 93. Although cross-reaction with C. ulcerans, a human/bovine species, was observed, the PCR assay used in this study may successfully be applied for the diagnosis of CL in goats and sheep as an alternative to conventional methods, owing to its advantages of specificity and speed.     

致羔羊脑炎粪肠球菌分离鉴定及毒力因子基因的PCR检测

从新疆部分地区羊场近年来发生的以脑炎为主要临床特征的病死羔羊体内分离出2株细菌,经鉴定为粪肠球菌。经链球菌A-G乳胶分型诊断液检测后确定为D群链球菌。该菌对小鼠和羔羊有较强的致病性,人工感染可引起小鼠的死亡,感染羔羊能够复制出与自然感染相同的临床症状和病理变化,并从感染发病和死亡羊的脑、心血、肝脏中再分离到相同细菌,分离菌对万古霉素、氟苯尼考、头孢噻肟钠、诺氟沙星和恩诺沙星敏感,根据GenBank中粪肠球菌主要毒力因子的基因序列设计引物进行PCR检测,结果显示,1株细菌明胶酶（GelE）、聚合物质（Asa1）、心内膜炎抗原（efaA）、胶原蛋白黏附素（Ace）、新的表面蛋白1（EF3314）等毒力因子基因阳性;另1株细菌溶血素激活因子（CylA）、表面蛋白（esp）、聚合物质（Asa1）、心内膜炎抗原（efaA）、胶原蛋白黏附素（Ace）、2种新的表面蛋白1和2（EF0591和EF3314）等毒力因子基因为阳性。     

Molecular discrimination of sheep and cattle isolates of Echinococcus granulosus by SSCP and conventional PCR in Turkey

Cystic echinococcosis (CE), caused by hydatid cysts, is a widespread and hazardous disease in humans and animals worldwide. The aim of the current study was to investigate the genetic characteristics of sheep and cattle isolates of Echinococcus granulosus obtained from eastern Turkey using Single Stranded Conformation Polymorphism (SSCP) analysis and conventional PCR method. A total of 54 isolates collected from Erzurum and Elazig provinces of east-Turkey were examined. The 31 of these were obtained from liver of sheep while 23 cattle isolates (12 of liver and 11 of lung) were tested. After the total genomic DNA isolation 12S rRNA gene of all isolates were examined by PCR for the aim of genetic characterization by conventional PCR and mitochondrial CO1 gene for SSCP analysis. The 12S rRNA-PCR yielded 254 bp of amplification product with all samples analyzed. Thus, these samples were identified as G1-G3 cluster (E. granulosus sensu stricto). At least two major single stranded bands were resolved for G1-G3 cluster and G5 in SSCP analysis. While the resolution of more than two additional single stranded bands in SSCP indicated the existence of G7 genotype. The SSCP analysis was identified the G5 and G7 while failed to G1 and G3. The present SSCP analysis classified all 54 cyst isolates from sheep and cattle as E. granulosus sensu stricto (G1-G3 cluster). However, some sequenced samples for G1 and G3 showed the same band patterns by SSCP.     

Comparison of bacterial cultivation, PCR, in situ hybridization and immunohistochemistry as tools for diagnosis of Haemophilus somnus pneumonia in cattle

The aim of the present study was to compare the potential of bacterial cultivation (BC), PCR, in situ hybridisation (ISH), and immunohistochemistry (IHC) in the diagnosis of Haemophilus somnus, when applied to pneumonic bovine tissue. Lungs from 65 field cases submitted for bacteriological examination were included in the study. The PCR-detection was performed on three different samples: plate-PCR (detection on plate washes after incubation of lung tissue on agar plates); swab-PCR (direct detection on a swab from the cut surface); and, whenever possible, a bronchus-PCR (direct detection on a swab from the main bronchus of the right cranial lung lobe). In order to examine the pathological significance of the findings, a histopathological examination of the cases was performed. H. somnus was detected by one or more techniques in 33 cases in total. By BC the bacterium was isolated from 10 cases, IHC and ISH were positive in 17 and 19 cases, and plate- and swab-PCR were positive in 21 and 29 cases, respectively. The bronchus-PCR was positive in 30 out of 61 cases examined. The PCR-technique was the most sensitive method, and as this technique is fast and relatively inexpensive, it should be considered as a supplementary tool in the diagnosis of H. somnus induced calf pneumonia.     

PrP genotype frequencies of Quebec sheep breeds determined by real-time PCR and molecular beacons

The allele and genotype frequencies of the prion protein gene (PrP), known to have an impact on scrapie susceptibility, were determined by real-time PCR for 500 Quebec purebred rams. Molecular beacons were very efficient in discriminating the 5 alleles investigated. Polymorphisms at coding positions 136, 154, and 171 of the PrP gene were analyzed using 3 separate real-time PCR reactions and a total of 7 molecular beacons. A total of 4 different alleles (ARQ, ARR, AHR, and VRQ) were observed at different frequencies among the 7 breeds of sheep investigated. Results show that more than 50% of the rams in every breed carried at least one ARR allele, which is considered the most resistant to scrapie. The susceptibility ARQ allele was also present in every breed and together with the ARR allele, they were the most frequent alleles found in Quebec rams. The VRQ allele associated with the highest susceptibility to scrapie occurred in 5 of the 7 breeds, although at low frequencies. Overall, the results indicate that the frequencies of PrP alleles and genotypes in common breeds of sheep in Quebec make it feasible to reduce scrapie risk by selective breeding.     

Pathological changes in the lungs and mammary glands of sheep and their relationship with maedi-visna infection.

Maedi-visna, a multisystemic disease of adult sheep, was first described in Spain in 1984. To get an idea of the seroprevalence of the disease locally and to estimate the number of seropositive animals with lesions, samples of blood, lungs and mammary glands were taken from 124 randomly selected sheep killed in the main slaughterhouse of Zaragoza. In the agar gel immunodiffusion test, 74 (59.7 per cent) of the sheep were positive and 50 were negative. Among the 74 seropositive animals, 19 (25.6 per cent) had no lesions in any organ, 12 (16.2 per cent) had lesions in the lungs only, 15 (20.2 per cent) had lesions in the mammary glands and 28 (37.8 per cent) had lesions in both organs. In the lungs hyperplasia of lymphoid follicles was more evident than an interstitial infiltrate but in the mammary glands this relationship was not observed. Even when the lesions occurred in both organs, they did not show the expected proportion in terms of either type or severity. Among the 50 seronegative sheep, eight (16 per cent) showed maedi-like lesions, formed exclusively by the hyperplasia of lymphoid follicles.     

多重PCR检测圈养牛、猪和羊源魏氏梭菌

采用多重PCR对圈养源魏氏梭菌的α，β，ε，ι毒素基因进行了检测，结果证实该方法具有很高的特异性。通过对山东德州、枣庄、泰安、蒙阴曾经流行过魏氏梭菌病的猪场、牛场和羊场的162个粪便样品进行检测，检出率为19．1％，均为A型。应用该方法鉴别魏氏梭菌血清型快速、简便，结果对于预防治疗均具有重要的指导作用。     

Assessment of Aspergillus fumigatus burden in lungs of intratracheally-challenged turkeys (Meleagris gallopavo) by quantitative PCR,galactomannan enzyme immunoassay,and quantitative culture

Aspergillus fumigatus remains a major respiratory pathogen in birds and treatment is still difficult. We challenged different groups of few-day-old turkeys via intratracheal aerosolisation with increasing concentrations (10⁵ up to 10⁸) of conidia using a MicroSprayer^® device. The fungal burden was assessed by real-time PCR, galactomannan dosage, CFU counting and histopathological evaluation in order to provide a comparison of these results within each inoculum groups. Significant mortality, occurring in the first 96 h after inoculation, was only observed at the highest inoculum dose. Culture counts, GM index and qPCR results on the one hand and inoculum size on the other hand appeared to be clearly correlated. The mean fungal burden detected by qPCR was 1.3 log₁₀ units higher than the mean values obtained by CFU measurement. The new model and the markers will be used to evaluate the efficacy of antifungal treatments that could be used in poultry farms.     

Diagnosis by culture and PCR of Salmonella Abortusovis infection under clinical conditions in aborting sheep in Switzerland

Since 2003 eleven Swiss sheep flocks were affected by abortion storms due to Salmonella abortusovis, an infection which had not been reported in this country for decades although cases of salmonellosis are notifiable in Switzerland. This raised doubts about the adequacy of the currently used diagnostic tools and the origin of this infection. Therefore, PCR was tested for its potential as a more rapid and more reliable method for diagnosing S. abortusovis infections under field conditions. Fecal and vaginal samples were collected at different times after abortion and PCR was used to detect bacterial DNA. Bacteria were isolated by conventional culture techniques. For determining their origin they were analyzed by pulsed field gel electrophoresis (PFGE) and compared to isolates from Germany and France. Sequencing of randomly selected amplicons allowed confirming the specificity of the result. PCR was more sensitive because it allowed detecting S. abortusovis DNA up to three months after infection even in samples that were negative by culture. Escherichia coli from the digestive tract of sheep could inhibit the growth of S. abortusovis in vitro suggesting that the lower sensitivity of diagnosis by bacterial culture may in part be due to growth inhibition of S. abortusovis by resident bacteria. Results of PFGE indicated that the Swiss strains were closely related among themselves but distinct from German and French strains suggesting the presence of an autochthonous infection.     

荧光定量PCR法检测饲料中的牛、羊成分

疯牛病是一种严重威胁畜牧业发展和人类生命安全的传染病，因此对于我们国家而言，急需建立一套严格，规范的检验标准，防止疫区的牛，羊肉骨粉通过直接或间接贸易进口，本文运用荧光定量PCR法，针对牛，羊两种不同动物的特异性基因序列设计带有不同荧光标记的特异性探针，对整个PCR过程采用实时监控，最终通过不同的荧光信息来鉴定饲料中的牛，羊成分。本方法能有效解决普通PCR的样品污染问题，具有特异性强，灵敏度高，重复性好等特点。     

Identification,typing, ecology and epidemiology of coagulase negative staphylococci associated with ruminants

Since phenotypic methods to identify coagulase negative staphylococci (CNS) from the milk of ruminants often yield unreliable results, methods for molecular identification based on gene sequencing or fingerprinting techniques have been developed. In addition to culture-based detection of isolates, culture-independent methods may be of interest. On the basis of molecular studies, the five CNS species commonly causing intramammary infections (IMI) are Staphylococcus chromogenes, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus simulans and Staphylococcus xylosus. Current knowledge suggests that S. chromogenes is a bovine-adapted species, with most cases of IMI due to this bacterium being opportunistic. S. haemolyticus also appears to be an opportunistic pathogen, but this bacterium occupies a variety of habitats, the importance of which as a source of IMI remains to be elucidated. S. xylosus appears to be a versatile species, but little is known of its epidemiology. S. epidermidis is considered to be a human-adapted species and most cases of IMI appear to arise from human sources, but the organism is capable of residing in other habitats. S. simulans typically causes contagious IMI, but opportunistic cases also occur and the ecology of this bacterium requires further study. Further studies of the ecology and epidemiology of CNS as a cause of IMI in cattle are required, along with careful attention to classification of these bacteria and the diseases they cause.     

Influence of management system on litter size, lamb growth, and carcass characteristics in sheep.

Three management systems (winter, spring, and late summer) distinguished by season of lambing and management practices were compared for litter size born and weaned and growth and carcass characteristics of lambs. Three sire breeds (Cheviot, Rambouillet, and Suffolk) and three dam breeds (Florida Native, Native-X, and Synthetic-X) were used in 698 matings over a 3-yr period. System affected (P < .01) litter size born. Spring lambing yielded more lambs (1.62) than winter (1.49) and late summer (1.12). The winter-born lambs were lighter but fatter (P < .05), and spring-born lambs were leaner with higher leg conformation and carcass quality scores. The late summer-born lambs were not different from spring-born lambs. Wethers had higher (P < .01) weights off test than ewe lambs (43.9 vs 42 kg) but had lower (P < .01) leg conformation scores, percentage kidney and pelvic fat, yield grade, and dressing percentages. Dam breed effects were significant (P < .05) for average preweaning daily gains with 249+/-5, 201+/-9, and 191+/-9 g for progeny of Native-X, Florida Native, and Synthetic-X, respectively. Single-born lambs had higher daily gains (P < .05) than twins in a preweaning period in all management systems and higher postweaning and lifetime daily gains for winter and spring management systems.     

Factors associated with ewe and lamb survival,and subsequent reproductive performance of sheep undergoing emergency caesarean section

Clinical records of all 212 ewes undergoing emergency caesarean surgery at a veterinary teaching hospital between January 2008 and December 2019 were evaluated retrospectively. Their age ranged from 1 to 10 years (median = 4 years), with German merino the predominant breed (48.1% of cases). The most frequently diagnosed indications were insufficient cervical dilatation (n = 94, 44.3%), uterine torsion (n = 50, 23.6%), foetopelvic disproportion (n = 31, 14.6%) and vaginal prolapse intra partum (n = 11, 5.2%). Fifty-four (25.5%) of the 212 ewes additionally suffered from one or more concurrent, pre-existing conditions. Overall ewe mortality until hospital discharge was 10.8% (23/212), and 3.8% (n = 6) for the 158 ewes without a history of concurrent disorders. Mortality during hospitalization increased to 31.5% (17/54) for those with pre-existing conditions. Total lamb mortality was 49.1% (173/352) until hospital discharge. Pre-existing conditions (p = .001) and the presence of post-surgical complications (p = .025) were identified as significant factors influencing dam mortality, while delayed presentation for veterinary attention with an observed duration of labour of >12 hr was identified as the most influential factor on total lamb mortality (p = .010). The presence of dead or emphysematous foetuses was not significant for ewe mortality. Follow-up information on further outcomes was available for 156 (82.5%) of the 189 discharged ewes. Eighty-nine animals (57.1%) were re-bred in the following season and achieved a 93.3% (83/89) pregnancy rate, while the remainder had either been slaughtered (n = 56, 35.9%), sold (n = 5, 3.2%) or had died of unknown causes (n = 3, 1.9%). The subsequent incidence of dystocia was 15.6% (n = 12) in the 77 ewes with available information on lambing ease. Adequate management of underlying conditions and timely intervention are important factors for best possible short-term outcomes. In the long term, the subsequent pregnancy rate was good and the incidence of subsequent dystocia was within the normal range.     

Prevalence of hydatidosis and fertility of hydatid cysts in sheep in Kirikkale,Turkey

This study was conducted in order to determine the prevalence and fertility of hydatid cysts in sheep in Kirikkale, Turkey. A total of 3.2% of 553 lambs and 50.9% of 1320 adults were found to be infected with hydatid cysts. The most common locations of cysts were in the liver and lung. Liver cysts tended to be more fertile than lung cysts (81.53% v. 76.47%), though the difference was not significant. The mean number of viable protoscoleces in the liver and lung cysts were 12,400 and 5,800, respectively. In general, 1-10 cysts were found in liver (51.8%) and lung (64.7%). The findings of this study indicate that hydatid cysts are common in sheep and the fertility rate of cysts is quite high. Sheep play an important role in the life cycle of Echinococcus granulosus in this region.     

Detection of Brucella species in the milk of infected cattle,sheep, goats and camels by PCR

One hundred and three milk samples were collected from 52 cows, 21 ewes, 18 goats and 12 camels. The animals tested positive to at least one of the following: (1) standard tube agglutination test (SAT); (2) Rose Bengal plate test (RBPT); (3) milk ring test (MRT). All milk samples were examined by culture and single-step polymerase chain reaction (PCR) techniques for detection of Brucella species. The PCR assay amplified Brucella-DNA from 29 bovine milk samples, 10 from sheep, 13 from goats and one from a camel. The direct culture method detected Brucella organisms from 24 samples of cows' milk, 12 from sheep, 10 from goats and failed to detect any Brucella organisms from camels' milk. PCR detected up to 100 colony forming units (CFU) of B. abortus per millilitre of milk in 100% of diluted milk samples, and 1000 CFU of B. melitensis from 70% of milk samples. Although the overall sensitivity of the PCR was higher than the culture method, it should be possible to increase the sensitivity to detect lower numbers of Brucella organisms in field samples. The speed and sensitivity of the PCR assay suggest that this technique could be useful for detection of Brucella organisms in bovine milk, as well as in sheep, goat, and camels milk.     

Seroprevalence of coxiellosis in cattle, sheep and people in the east of Turkey

Serum samples collected randomly from 416 cattle in 48 herds, and 411 sheep in 47 flocks, in eight different locations in the east of Turkey between June and December 1998, were examined by indirect fluorescent antibody test (IFAT) to determine the prevalence of Q fever. The age, sex, breed, tick control and abortion history of the animals were also recorded. In addition, 102 serum samples were collected from apparently healthy people who were at risk of contracting the disease, such as farmers, veterinarians, abattoir and laboratory workers, and veterinary students. Seropositivity was observed in 5-8 per cent (24/416) of the cattle in 17 (35-4 per cent) of the herds and in 10-5 per cent (43/411) of the sheep in 21 (44-7 per cent) of the flocks. Eight of the 102 people were seropositive, with the highest prevalence (12-0 per cent) in farmers and abattoir workers. All the seropositive farmers had seropositive animals. None of the laboratory workers or veterinary students was seropositive.