Detection of Hepatitis E Virus (HEV) Among Ruminant Animals.

BACKGROUND: HEV is endemic in several Middle Eastern countries including Saudi Arabia, which hosts the annual pilgrimage for Muslims from around the world. One of the Hajj rituals is the sacrifice of animals, including camels, cows, goats, and sheep. HEV Zoonosis is established in swine and other suspected species, including deer, rabbits, dromedary, and Bactrian camels. HEV was identified in small, domesticized animals like goats, cows, sheep, and horses. We previously investigated HEV seroprevalence in Camels. This study aimed to evaluate HEV seroprevalence in other highly consumed ruminants in Saudi Arabia, namely cows, sheep, and goats. METHODS: Sera from cows (n = 47), goats (n = 56), and sheep (n = 67) were analyzed for the presence of HEV-IgG by using in-house developed ELISA assays. RESULTS: The highest seroprevalence was found in sheep (62.7%), followed by cows (38.3%), and then goats (14.3%), with a p-value of < 0.001. No other demographic characteristics of the animals were significantly correlated with the HEV seroprevalence. CONCLUSIONS: This study provides baseline data as the first study on the seroprevalence of HEV in ruminant animals in Saudi Arabia. The high seroprevalence found in sheep and cows must be further investigated for the potential zoonotic HEV transmission to humans. Further studies are needed to investigate the active viremia in these animal species through nucleic acid detection and sequencing to provide data on the circulating HEV genotypes among the targeted animal species. The detection of HEV in different animal products, such as milk, liver, and others, also remains an important study area to consider.

Evaluation of Oxidative Stress, Thyroid Hormones, Trace Elements and Some Biochemical Markers in Goats Naturally Infected with Theileria ovis.

BACKGROUND: Theileriosis is a tick-borne disease caused by protozoon species in the Theileria genus of the Theileriidae family. The biochemical changes induced by infection are considered to be an important understanding of the pathophysiology of caprine theileriosis. In this study, it was aimed to determine oxidative stress, thyroid hormones, trace elements, and biochemical parameters in theileriosis infection. MATERIALS AND METHODS: A sample of 14 goat was used for this purpose, of which 7 were healthy and 7 were infected with Theileria ovis. Theileria infection was diagnosed from the polymerase chain reaction (PCR). Sera from blood samples was tested for total antioxidant capacity (TAC), total oxidant capacity (TOC), oxidative stress index (OSI), alanine aminotransferase (ALT), aspartate transaminase (AST), gamma glutamyl transferase (GGT), total protein, albumin, triglyceride, cholesterol, high density lipoprotein (HDL), low density lipoprotein (LDL), urea, blood urea nitrogen (BUN), creatinine, triiodothyronine (T3), thyroxine (T4), copper (Cu), zinc (Zn), cobalt (Co), manganese (Mn), selenium (Se), iron (Fe). RESULT: TOC, OSI, AST, ALT and GGT values were higher in the patient group than in the healthy group (P < 0.05). On the other hand, there were decreases in TAC, T3, T4, total protein, albumin, creatinine, Cu, Zn, Se, and Co values (P < 0.05). However, there was not found to be a statistical difference between the healthy and patient groups in terms of triglyceride, cholesterol, HDL, LDL, urea, BUN, Mn, and Fe values (P > 0.05). CONCLUSIONS: It can be stated that oxidative stress is a complication of caprine theileriosis and it may be accompanied with hypothyroidism and deficits in trace minerals.

Validation of a reference interval for symmetric dimethylarginine in healthy goats and its comparison to values in goats with obstructive urolithiasis.

BACKGROUND: Symmetric dimethylarginine (SDMA), a sensitive biomarker for detecting renal injury, has not been characterized in goats. Obstructive urolithiasis (OU) is the most common urinary tract disease in male small ruminants. HYPOTHESIS/OBJECTIVE: Establish an SDMA reference interval (RI) in healthy adult goats and describe SDMA concentrations in goats with OU. We hypothesize that the SDMA RI in healthy adult goats will be similar to that of other adult veterinary species and that SDMA can be utilized to assess the renal function of goats experiencing OU. ANIMALS: Fifty-five healthy adult male and female goats from a university herd were enrolled for SDMA RI development. Twenty male and female goats from a university herd were enrolled for validation of the SDMA RI established. Thirteen male goats diagnosed with OU were enrolled. METHODS: Clinical trial. Serum samples for all animals enrolled were collected and analyzed for SDMA using an immunoassay (IDEXX Laboratories, Inc); goats with OU had additional blood work analyzed (PCV, total solids, and serum biochemistry). Symmetric dimethylarginine and other values in goats with OU were analyzed and compared at specific time points. RESULTS: The SDMA RI for healthy, adult goats is 8.03 µg/dL (90% CI 4.81-11.04) to 25.93 µg/dL (90% CI 22.88-28.97). There was no correlation identified between serum creatinine and SDMA in goats with OU. CONCLUSIONS AND CLINICAL IMPORTANCE: The SDMA RI for adult goats is higher than in other adult large animal species. Use of SDMA in goats with OU is not useful in assessing their renal function.

Herd-level true seroprevalence of caseous lymphadenitis and paratuberculosis in the goat population of Poland.

A large-scale study was carried out in the Polish goat population in 2014-2021 to determine the herd-level true seroprevalence (HTP) of caseous lymphadenitis (CLA) caused by Corynebacterium pseudotuberculosis (Cp) and paratuberculosis (PTB) caused by Mycobacterium avium ssp. paratuberculosis (Map). Two-stage cluster sampling was applied to herds counting at least 20 adult goats (aged >1 year) and in each herd all males and 10-13 females were tested. At least one seropositive goat regardless of its sex was necessary to consider the herd as infected. HTP was estimated using the Bayesian approach with the Gibbs sampler in the EpiTools and reported as the median and 95 % credibility interval (95 % CrI). A total of 1282 adult goats from 86 herds were serologically tested using two commercial ELISAs (Cp-ELISA and Map-ELISA). At least 1 seropositive result of Cp-ELISA and Map-ELISA was obtained in 73/86 herds (84.9 %) and 40/86 herds (46.5 %), respectively. HTP of CLA was estimated at 73.3 % (95 % CrI: 65.0 %, 80.4 %) and HTP of PTB was estimated at 42.9 % (95 % CrI: 25.8 %, 58.0 %). There was a significant positive association between the occurrence of CLA and PTB in the herds (odds ratio 6.0, 95 % confidence interval: 1.2, 28.8; p = 0.010). Probability of the seropositive result for PTB was also significantly higher in Cp-seropositive goats than in Cp-seronegative goats (odds ratio 3.9, 95 % confidence interval: 2.4, 6.3; p < 0.001) which could indicate either a higher risk of co-infection or a higher rate of false positive results for PTB in Cp-positive goats. To investigate this issue, optical densities obtained in Map-ELISA were compared between Cp-positive and Cp-negative goats and results of Map-ELISA were adjusted accordingly. Map-negative sera from Cp-positive goats turned out to have significantly higher optical densities than Map-negative sera from Cp-negative goats (p < 0.001). After the adjustment, the herd-level apparent seroprevalence of PTB was 41.9 % (36/86 herds) so it still fell within the 95 % CrI of HTP of PTB calculated before the adjustment. Concluding, CLA appears to be widespread in the Polish goat population. In many of them it may be subclinical at the moment, however will likely emerge in the future as the disease follows cyclic pattern in Poland. On the other hand, given the total lack of clinical PTB in Polish goats, an explanation for a high HTP of PTB remains unclear and warrants further studies using tests of higher analytical specificity than ELISA.

Molecular prevalence, associated risk factors and phylogenetic evaluation of Theileria lestoquardi in the blood samples of small ruminants.

Raising small ruminants is the main source of income for farmers in Pakistan especially in rural areas of Dera Ghazi Khan in Punjab. Despite having large sheep population, the prevalence of intra-erythrocytic protozoa, Theileria (T.) lestoquardi, has never been reported from this area. This study was conducted to fill this knowledge gap and 333 blood samples of apparently healthy small ruminants (168 sheep and 165 goats) along with their epidemiological data were collected from Dera Ghazi Khan district during August till November 2022. The polymerase chain reaction (PCR) analysis amplified a 785 base pair amplicon specific for the Merozoite surface antigen (ms 1-2) gene of T. lestoquardi in 2 out of the 168 (3.3%) sheep blood samples, while no goat blood sample out of 165 was found to be infected with T. lestoquardi. DNA sequencing confirmed the presence of Theileria lestoquardi in both samples and phylogenetic analysis revealed that these amplicon resembled the partial ms 1-2 gene sequences detected in small ruminants from Pakistan, India Iran and Egypt. All the studied epidemiological factors (age, sex, breed, size of herd, dogs with herd, composition of herd, size of herd and Tick burden on sheep) were not found associated with the prevalence of T. lestoquardi. In conclusion, this study reports a low prevalence of T. lestoquardi infection in the Dera Ghazi Khan District of Punjab, Pakistan. The data generated from this work will help pave the way for the prophylactic detection and control of ovine and caprine theileriosis in the region.

Serological investigation and genotyping of Mycobacterium avium subsp. paratuberculosis in sheep and goats in Inner Mongolia, China.

Paratuberculosis a contagious and chronic disease in domestic and wild ruminants, is caused by Mycobacterium avium subspecies paratuberculosis (MAP). Typical clinical signs include intractable diarrhea, progressive emaciation, proliferative enteropathy, and mesenteric lymphadenitis. Paratuberculosis is endemic to many parts of the world and responsible for considerable economic losses. In this study, different types of paratuberculosis and MAP in sheep and goats were investigated in Inner Mongolia, a northern province in China contiguous with two countries and eight other provinces. A total of 4434 serum samples were collected from six cities in the western, central, and eastern regions of Inner Mongolia and analyzed using the ELISA test. In addition, tissue samples were collected from seven animals that were suspected to be infected with MAP. Finally, these tissues samples were analyzed by histopathological examination followed by polymerase chain reaction (PCR), IS1311 PCR-restriction enzyme analysis (PCR-REA), and a sequence analysis of five genes. Among all 4434 ruminant serum samples collected from the six cities in the western, central, and eastern regions of Inner Mongolia, 7.60% (337/4434) measured positive for the MAP antibody. The proportions of positive MAP antibody results for serum samples collected in the western, central, and eastern regions were 5.10% (105/2058), 6.63% (85/1282), and 13.44% (147/1094), respectively. For the seven suspected infected animals selected from the herd with the highest rate of positivity, the gross pathology and histopathology of the necropsied animals were found to be consistent with the pathological features of paratuberculosis. The PCR analysis further confirmed the diagnosis of paratuberculosis. The rest of the results demonstrated that herds of sheep and goats in Inner Mongolia were infected with both MAP type II and type III. To the best of our knowledge, this is the first study of the two subtypes of MAP strains in sheep and goats in Inner Mongolia.

Caprine Arthritis Encephalitis Virus Is Associated with Renal Lesions.

Caprine arthritis encephalitis virus (CAEV) is a monocyte/macrophage-tropic lentivirus that primarily infects goats resulting in a well-recognized set of chronic inflammatory syndromes focused on the joint synovium, tissues of the central nervous system, pulmonary interstitium and mammary gland. Clinically affected animals generally manifest with one or more of these classic CAEV-associated tissue lesions; however, CAEV-associated renal inflammation in goats has not been reported in the peer-reviewed literature. Here we describe six goats with chronic, multisystemic CAEV infections in conjunction with CAEV-associated renal lesions. One of the animals had CAEV antigen-associated thrombotic arteritis resulting in infarction of both the kidney and heart. These goats had microscopic evidence of inflammatory renal injury (interstitial nephritis) with detectable renal immunolabeling for CAEV antigen in three of six animals and amplifiable proviral sequences consistent with CAEV in all six animals. Cardiac lesions (vascular, myocardial or endocardial) were also identified in four of six animals. Within the viral promoter (U3) region, known transcription factor binding sites (TFBSs) were generally conserved, although one viral isolate had a duplication of the U3 A region encoding a second gamma-activated site (GAS). Despite the TFBS conservation, the isolates demonstrated a degree of phylogenetic diversity. At present, the clinical consequence of CAEV-associated renal injury is not clear.

Sero-epidemiology and associated risk factors of brucellosis among sheep and goat population in the south western Nepal: a comparative study.

BACKGROUND: Brucellosis is a zoonotic disease caused by Brucella spp. In Nepal, the presence of brucellosis in small ruminants, namely sheep and goats, has impacted farmers' livelihood and the food safety of consumers. A cross-sectional study was conducted in Rupandehi district of Nepal during January to March 2020 to investigate the seroepidemiology and associated risk factors of brucellosis in the sheep and goat population. Altogether, 19 sheep and 60 goat farms in the district were visited. Owners were interviewed to get information on animals, including their management and movement patterns. Three hundred fifty-seven samples (80 sheep and 277 goat samples) were collected proportionately based on farm sizes. Each serum sample was tested with Rose Bengal Test and ELISA to estimate the seropositivity of brucellosis. Logistic regression was carried out to calculate corresponding odds ratios of each variable associated with detection of brucellosis. RESULTS: At the farm level, 31.6% (6/19; 95% CI: 12, 54%) of sheep farms and 3.3% (2/60, 95% CI: 0.9, 11.4%) of goat farms were seropositive to brucellosis. Out of 80 sheep serum samples, 12 (15%; 95% CI: 8.79-24.41%) and out of 277 goat serum samples, three (1.1%; 95% CI: 0.37-3.14%) were seropositive to brucellosis. Age greater than 1.5 years (OR = 5.56, 95% CI: 1.39, 29.38; p = 0.02) and herd size of greater than 100 (OR = 4.74, 95% CI: 1.23, 20.32, p = 0.03) were identified as significant risk factors for seropositivity of brucellosis in the sheep population. While in the goat population, none of the variables was identified as a significant risk factor. CONCLUSION: The study provides evidence that the older sheep and the sheep from the large herds were at higher risk of brucellosis. A control program should be put in place immediately in the sheep population because they may transmit infections to other livestock as they were regularly moved for grazing and selling purposes. Also, strict biosecurity measures should be implemented among pastoralists to prevent brucellosis transmission in them. We suggest further one health-based study to reveal the transmission dynamics of brucellosis between animals and humans.

Hematologic Conditions of Small Ruminants.

Anemia is a clinically important syndrome in small ruminants. Anemia can be divided into regenerative and nonregenerative forms. Differentials for regenerative anemia include hemorrhage owing to gastrointestinal or external parasitism or hemostatic disorders, and hemolysis owing to infectious, osmotic, toxic, and nutritional causes. Differentials for nonregenerative anemia include inflammatory and chronic diseases, renal failure, pancytopenia, copper deficiency, and heavy metal toxicosis. Iron deficiency anemia can be caused by chronic gastrointestinal and external hemorrhage or nutritional deficiency and may be mildly regenerative or nonregenerative. Appropriate diagnostic tests are described along with treatments, including blood transfusion, parasite control, and prevention.

Development and evaluation of recombinant GRA8 protein for the serodiagnosis of Toxoplasma gondii infection in goats.

BACKGROUND: The development of sensitive and specific methods for detecting Toxoplasma gondii infection is critical for preventing and controlling toxoplasmosis in humans and other animals. Recently, various recombinant proteins have been used in serological tests for diagnosing toxoplasmosis. The production of these antigens is associated with live tachyzoites obtained from cell cultures or laboratory animals for genomic extraction to amplify target genes. Synthetic genes have gained a key role in recombinant protein production. For the first time, we demonstrated the production of the recombinant protein of the T. gondii dense granular antigen 8 (TgGRA8) gene based on commercial gene synthesis. Recombinant TgGRA8 plasmids were successfully expressed in an Escherichia coli system. The recombinant protein was affinity-purified and characterized via sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Furthermore, the diagnostic potential of the recombinant protein was assessed using 306 field serum samples from goats via indirect enzyme-linked immunosorbent assay (iELISA) and the latex agglutination test (LAT). RESULTS: Western blotting using known positive serum samples from goats identified a single antigen at the expected molecular weight of TgGRA8 (27 kDa). iELISA illustrated that 15.40% of goat samples were positive for T. gondii-specific IgG antibodies. In addition, TgGRA8 provided high sensitivity and specificity, with significant concordance (91.83) and kappa values (0.69) compared with the results obtained using LAT. CONCLUSION: Our findings highlight the production of a recombinant protein from a synthetic TgGRA8 gene and the ability to detect T. gondii infection in field samples. The sensitivity and specificity of TgGRA8 demonstrated that this protein could be a good serological marker for detecting specific IgG in goat sera.

Prevalence, distribution and risk factors for brucellosis infection in goat farms in Ningxiang, China.

BACKGROUND: In south China, goats are the major source of Brucellosis for human infection. However, there are few studies on the prevalence of and risk factors for goat brucellosis in south China. In this study, we conducted a cross-sectional study to investigate the herd prevalence, spatial distribution and relevant risk factors for goat brucellosis in Ningxiang county, south China. Commercial goat farms (n = 457) were randomly selected, and their disease status was ascertained by testing serum samples of chosen individuals using the Rose Bengal Test (screening test) and the Serum Agglutination Test (confirmatory test) in series. A farm with at least two positive individuals was defined as a case farm. Standardized questionnaires were used to collect information on management and hygiene practices in farms. A logistic model with a binomial outcome was built to identify risk factors for being seropositive. RESULTS: The true herd prevalence in commercial goat farms was 4.5% (95%CI: 0.2%-12.2%) and the townships in the centre of the county had higher herd prevalence. The risk factors associated with seropositive on local goat farms include "Introduction in the past 12 months" (OR= 61, 95%CI: 16-333), "Improperly disposal of the sick or dead goats" (OR= 33, 95%CI: 5-341) and "Poor hygiene in lambing pen" (OR= 25, 95%CI: 5-192). CONCLUSIONS: These findings will aid in the development of control strategies of Brucellosis in south China and risk factors identified in this study should be taken into consideration when designing a control strategy.

Sequential haematological and serum biochemical changes in Black Bengal goats infected with a local isolate of peste des petits ruminants virus from Bangladesh.

BACKGROUND: Knowledge of sequential changes in haematobiochemical parameters of infected animals helps in the formulation of appropriate supportive therapy. OBJECTIVE: We investigated the sequential haematological and biochemical changes in peste des petits ruminants (PPR)-infected Black Bengal goats. METHODS: Goats were either infected with PPR virus (PPRV; n = 8) or sham infected with sterile phosphate-buffered saline (n = 4) via the intranasal route. Blood and sera were collected from both groups at different days post-infection (dpi) and analysed. Goats were sacrificed at different dpi and the amount of PPRV RNA in different tissues was quantified by real-time RT-PCR. RESULTS: The PPRV-infected goats showed mild depression and scanty nasal secretions starting at 4 dpi which became severe with high fever (106°F), dyspnoea, stomatitis, profuse orinasal discharge and diarrhoea at 9-13 dpi. PPRV RNA was detected in different tissues of infected goats. Severe lymphocytic leukopenia (at 18 dpi) was observed in infected goats. Total protein and albumin decreased in infected goats starting at 10 dpi. An elevated level of enzymes (alkaline phosphatase, creatine kinase, aspartate transaminase and alanine transaminase) and metabolites (blood urea nitrogen and urea B) were found in infected goats starting at 7-10 dpi, suggesting damages in the liver and kidneys. PPR-infected goats showed elevated sodium and chloride ions starting at 7 dpi. The majority of infected goats were seroconverted by 14 dpi. CONCLUSIONS: Anti-diarrheal agents, aqua solutions and other medicine to support liver and kidney functions could be considered as supportive therapy against PPRV infection.

Isolation and Cultivation of a New Isolate of BTV-25 and Presumptive Evidence for a Potential Persistent Infection in Healthy Goats.

Recently, several so-called "atypical" Bluetongue virus (BTV) serotypes were discovered, including BTV-25 (Toggenburg virus), in Switzerland. Most "atypical" BTV were identified in small ruminants without clinical signs. In 2018, two goats from a holding in Germany tested positive for BTV-25 genome by RT-qPCR prior to export. After experimental inoculation of the two goats with the BTV-25 positive field blood samples for generation of reference materials, viremia could be observed in one animal. For the first time, the BTV-25-related virus was isolated in cell culture from EDTA-blood and the full genome of isolate "BTV-25-GER2018" could be generated. BTV-25-GER2018 was only incompletely neutralized by ELISA-positive sera. We could monitor the BTV-25 occurrence in the respective affected goat flock of approximately 120 goats over several years. EDTA blood samples were screened with RT-qPCR using a newly developed BTV-25 specific assay. For serological surveillance, serum samples were screened using a commercial cELISA. BTV-25-GER2018 was detected over 4.5 years in the goat flock with intermittent PCR-positivity in some animals, and with or without concomitantly detected antibodies since 2015. We could demonstrate the viral persistence of BTV-25-GER2018 in goats for up to 4.5 years, and the first BTV-25 isolate is now available for further characterization.

Evaluation of inflammatory biomarkers in goats naturally infected with Babesia ovis.

This study was designed to evaluate the effects of Babesia ovis infection on concentrations of some essential acute phase proteins (APPs) including albumin, fibrinogen, serum amyloid A, haptoglobin, and ceruloplasmin as well as total, protein-binding, and lipid-binding sialic acids (TSA, PBSA, and LBSA) and two crucial cytokines including interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α). Some hematological parameters also were evaluated. Furthermore, any probable correlation among the APPs, SAs, IFN-γ, and TNF-α was calculated. A total of 420 Marghoz and Raeini goats with the ages of 1-3 years old from the north and northwest of Iran were examined, and 17 goats confirmed to be infected with B. ovis by both routine microscopic examination of blood films and molecular assays. As the control, 17 healthy goats were included. The results revealed a significant decrease (P < 0.05) in erythrocyte count, hemoglobin level, and pack cell volume as well as a nonsignificant increase in white blood cell count in the diseased animals compared with the control. Additionally, all the APPs, SAs, and cytokines were remarkably higher in the infected animals than the uninfected ones, except for albumin, which was significantly lower. Moreover, a strong and positive correlation was detected among the parameters mentioned above, except for albumin, which was inversely correlated with the other parameters. In conclusion, B. ovis infection is associated with the induction of severe inflammatory reactions in goats, and both SA and APP are significantly involved in the pathophysiology of the disease.

Gastrointestinal nematode infections in goats: differences between strongyle faecal egg counts and specific antibody responses to Teladorsagia circumcincta in Nera di Verzasca and Alpine goats.

Strongylida are gastrointestinal nematodes (GIN) of greatest importance in small ruminants throughout the world. Differences in resistance and resilience to GIN among goat breeds were reported. This study aims to investigate the mechanism underlying the breed-associated differences using a cosmopolitan (Alpine, AB) and an autochthonous (Nera di Verzasca, NV) goat breed. At first, fifteen goats from the same herd (NV = 7, AB = 8) at day 0 were infected with infective larvae (L3) of mixed GIN. From the 15th day post-infection (DPI), individual parasite egg excretion (faecal egg counts, FEC) was performed on all goats, once per week, until the 63rd DPI. Afterwards, in goats under field conditions (30 AB and 30 NV reared on the same farm), individual faecal and blood samples were collected; FEC-specific antibody and PCV levels were explored. In goats with experimental GIN infection, mean eggs per gram of faeces (EPG) values were consistently lower in NV goats. In goats with natural GIN infection, EPG and prevalence values showed high variability in both breeds; among individual variables, breed had a significant influence on EPG. Further, PCV and anti-T. circumcincta IgA levels were influenced by the breed. Lower PCV values were also associated with higher strongyle EPG in AB goats, and anti-T. circumcincta IgA levels were influenced by both strongyle EPG and breed, with IgA levels being higher in AB vs. NV goats and positively associated with EPG. Neither EPG nor breed had any influence on IgE levels. Both studies on experimental and natural infection confirmed that goats of NV are more resistant to infection with gastrointestinal nematodes.

First Report on Molecular Characterization of Anaplasmosis in Small Ruminants in Pakistan.

Anaplasmosis is caused by a Gram-negative obligate intracellular bacterium of the genus Anaplasma with the pathogen having a zoonotic impact. The study aimed to estimate the prevalence of anaplasmosis in Pakistan, to unravel the association of potential risk factors, and to investigate the effect on hematological parameters in affected small ruminants. A total of 150 (n = 75 sheep; n = 75 goats) blood samples were initially screened microscopically and then subjected to PCR targeting the amplification of the 16S rRNA gene fragment of Anaplasma. The PCR-based positive samples were then processed for sequencing. Statistical analysis regarding risk factors was performed using R software. The study revealed an overall 29.33% (44/150) prevalence of anaplasmosis in small ruminants. Sheep had higher (P > 0.05) prevalence (32%) as compared to goats (25.30%). The final statistical model resulting from backward elimination showed only tick infestation as a significant predictor of infection status. The phylogenetic analysis of 16S rRNA gene of Anaplasma spp. revealed 9 study isolates clustered together and showed a close resemblance (99%) with Anaplasma ovis isolate (DQ837600) from Hungary. One of the isolates showed (99%) similarity with the isolate of Anaplasma marginale (MH155594) from Iraq. Furthermore, the hematological parameters pack cell volume, red blood cells, hemoglobin, white blood cells, granulocytes, monocytes, lymphocytes, and platelet count were decreased in Anaplasma-positive animals. This is the first study at the molecular level to characterize Anaplasma spp. in small ruminants of Pakistan, and it will be useful in developing control strategies for anaplasmosis.

Short communication: Associations between nonesterified fatty acids, ß-hydroxybutyrate, and glucose in periparturient dairy goats.

The objective of the present study was to use longitudinal data to examine the relationships between blood concentrations of nonesterified fatty acids (NEFA), ß-hydroxybutyrate (BHB), and glucose during the transition period in dairy goats. Weekly blood samples were collected from Saanen goats from a commercial herd in Australia [1-7 yr; body weight 70 ± 16.0 kg; body condition score 2.5 ± 0.3; and daily milk yield 2.4 ± 0.73 L/d; all mean ± standard deviation (SD)]. The weekly prevalence of goats above hyperketonemic levels (BHB ≥0.8 mmol/L) was approximately 6 times greater postpartum than antepartum. As well, of the 935 goats sampled antepartum, 50 (5%) had at least 1 hyperketonemic event, and 823 (88%) had at least 1 event of NEFA above the threshold (≥0.3 mmol/L). Of 847 goats tested postpartum, 258 (30%) had at least 1 hyperketonemic event, and 690 goats (81%) had at least 1 event of NEFA above the threshold (≥ 0.7 mmol/L). Substantial variation was found when analyzing the mean days of maximum NEFA and maximum BHB concentrations antepartum (-11 ± 6.6 and -14 ± 7.2 d, respectively, mean ± SD) and postpartum (14 ± 6.6 and 9 ± 6.8 d, respectively, mean ± SD). We observed moderate to strong relationships between NEFA and BHB concentrations (r = 0.66) and between NEFA and glucose concentrations (r = -0.46) throughout the transition period. Our results suggested that 3 to 16 d in milk is the best sampling window for monitoring hyperketonemia in dairy goats, and that results from simultaneous BHB and glucose tests provide an improved indication of the fat mobilization and energy status of the herd when measured close to this timeframe.

Hematological prediction study of peritonitis following laparotomy in goats.

Surgical trauma to the abdominal wall and peritoneum during celiotomy is expected to cause postoperative inflammation. However, complications after abdominal surgery are hardly detected in the early stage. Hematological analysis of blood has been considered beneficial in disease diagnosis and prognosis. This study aimed to evaluate the hematological parameters predicting peritonitis in goats and to determine the post-surgery day that hematology is significant. Six apparently healthy West African Dwarf goats were included in this study. After 2 weeks of acclimatization, blood samples were obtained daily for 3 days for hematological analyses, which served as the baseline data. The right flanks of the animals were aseptically prepared routinely for exploratory laparotomy. Restraint and anesthesia were achieved using xylazine and lignocaine using an inverted "L" block technique. Laparotomy was performed, and the incision was left for 20 min and then closed routinely. Blood samples were collected for hemogram 24 hr postoperatively and daily for 7 days. Based on the post-surgery hematology results, relative neutrophil (P=0.015) and lymphocyte (P=0.006) counts significantly increased and decreased on day 5 respectively. Significant differences were also observed for red blood cell count, hemoglobin concentration, and packed cell volume on days 5, 6, and 7 respectively. It could therefore be concluded that the diagnostic result for hematology post-laparotomy can be obtained on the fifth and sixth day.

A highly sensitive semi-nested real-time PCR utilizing oligospermine-conjugated degenerate primers for the detection of diverse strains of small ruminant lentiviruses.

Small ruminant lentiviruses (SRLVs) are highly diverse retroviruses infecting sheep and goats. Although PCR-based testing is being utilized for diagnostics, its application is hampered by various factors. These include, among others, the exceptionally high genetic variability of SRLVs, as well as the low number of infected blood monocytes. For this reason, a highly sensitive and specific semi-nested real-time PCR for proviral DNA detection and quantification was developed. The method is innovative in that a) its design is based on selecting the preferred codon usage in the targeted conserved genomic regions and b) oligospermine-conjugated degenerate primers with increased Tm were utilized. Modifications permitted primer/template duplex formation in the cases of mismatches due to sporadic nucleotide polymorphisms in a number of variant SRLV strains and consequently, the detection of highly diverse SRLV strains. The potential loss of analytical sensitivity and specificity was counterbalanced by including a semi-nested step in combination with LNA probes. An in silico procedure for the evaluation of hybridization efficiency of the designed oligonucleotides to all known targeted variants was also implemented. The method presents a linear range of quantification over a 3-log10 range and a limit of detection of 3.9 proviral dsDNA copies per reaction. Its diagnostic performance was evaluated by testing field samples from seropositive and seronegative animals, followed by phylogenetic analysis of the strains detected. To further increase the diagnostic sensitivity, a DNA extraction protocol for blood leukocytes was developed and evaluated. A minimum of 500 ng input DNA is recommended for PCR-based detection of SRLV proviral DNA, given the low numbers of infected blood monocytes. The developed methodology may serve as a useful tool, which can be adjusted for the quantitative detection of viruses exhibiting high genetic variability.

Serological evidence of Coxiella burnetii infection in slaughtered sheep and goats at Kumasi Abattoir, Ghana.

Q fever, caused by Coxiella burnetii, is an important zoonosis worldwide. Q fever is documented in many parts of the world; however, information on the disease in Ghana is scanty. This study was therefore conducted to provide evidence of exposure of sheep and goats slaughtered at the Kumasi Abattoir to Coxiella burnetii. A total of 350 serum samples collected from 175 sheep and 175 goats were analyzed for the presence of C. burnetii antibodies using a commercial ELISA kit (ID Vet). Results of the study established a seroprevalence of 28.57% in goats, 16.57% in sheep and an overall seroprevalence of 22.29% in sheep and goats; 20.57% for male sheep, 23.86% for female sheep, 26.44% for male goats and 30.68% for female goats. Results showed that goats are more at risk to the infection than sheep however sex is not a risk factor. This study confirms the existence of Q fever in sheep and goats in Ghana hence, the disease should be considered as a public health risk to workers at the abattoir and other stakeholders in the sheep and goat production chain.