Introduction: The extremely high genetic variation and the continuously emerging variants of foot-and-mouth disease virus (FMDV) of Southern African Territory (SAT) serotypes including SAT1, SAT2, and SAT3 make it necessary to develop a new RT-PCR for general use for monitoring viruses based on the updated genome information. Material and Methods: A FMDV SAT-D8 one-step RT-PCR was established based on the 1D2A2B genes of the SAT serotype viruses with a multiplex primer set. FMDV A, O, C, and Asia 1 serotypes, other vesicular disease viruses, inactivated SAT viruses, and 125 bovine, ovine, caprine and porcine tissue samples collected from the Chinese mainland were included for evaluating the assay. Results: The new RT-PCR was proven to be specific without cross-reactions with Eurasian FMDV, swine vesicular disease virus (SVDV), Seneca valley virus (SVV), or other common viral pathogens of cattle, sheep, goat, and pig. An around 257 bp-sized amplicon clearly appeared when the inactivated SAT viruses were detected. However, all 125 samples collected from FMDV-susceptible animals from the Chinese mainland which has not known SAT epidemics showed negative results. Conclusions: A FMDV SAT-D8 one-step RT-PCR is a promising method for primary screening for FMDV SAT serotypes.

Introduction: Chlamydia psittaci is a gram-negative obligate intracellular pathogen of birds. Poultry infections lead to economic losses and can be transmitted to humans. No vaccine is available and the bacterium-host cell interaction is not completely understood. Replicating bacteria cause pneumonia, but C. psittaci can also be non-replicating and persistent inside the cytoplasm of avian cells. RT-qPCR provides insight into the molecular pathogenesis of both active replicating and persistent Chlamydia psittaci in birds, but requires identification of stably expressed reference genes to avoid biases. Material and Methods: We investigated the expression stability of 10 C. psittaci candidate reference genes for gene expression analysis during normal growth and penicillin-induced persistence. C. psittaci Cal10 was cultured in HeLa229 and RNA was extracted. The expression level of each candidate was examined by RT-qPCR and Cq values were analysed using geNorm. Results: The genes tyrS, gidA, radA, and 16S rRNA ranked among the most stably expressed. The final selected reference genes differed according to the bacterial growth status (normal growth versus persistent status), and the time points selected during the duration of the normal chlamydial developmental cycle. Conclusion: The study data show the importance of systematic validation of reference genes to confirm their stability within the strains and under the conditions selected.

Objective: The influence of intramammary infection (IMI) and types of bacteria was assessed on somatic cell count (SCC) in dairy cows milk with respect to breed, age, parity, stage of lactation, milk production, and mammary quarter location. Materials and methods: After recording data in a structured questionnaire, 360 samples of quar¬ter milk were collected. The samples were subjected to SCC and isolation and identification of bacteria. The data were analyzed to find out the significant influence of independent factors on SCC and IMI. Results: The infected quarters had a significantly higher mean SCC (210.52 × 103 cells/ml) compared to uninfected ones (32.72 × 103 cells/ml). The mean SCC was the highest for IMI with Enterobacter spp. (338.00 × 103 cells/ml) followed by Bacillus spp. (319.20 × 103 cells/ml), coagulase-negative Staphylococci (CNS) (268.17 × 103 cells/ml), Staphylococcus aureus (218.31 × 103 cells/ ml), and Escherichia coli (200.75 × 103 cells/ml) and the lowest for Pseudomonas aeruginosa (66.33 × 103 cells/ml). Milk of rear quarters had a significantly higher SCC than the front quarters. SCC increased with increasing age, parity, and lactation stage regardless of whether cows are infected or not. The IMI was more prevalent in rear quarters (42.2%) and cows at early (≤7 days) lactation (100.0%). Cows having a parity of ≥5 and crossbred and high yielding (>5 l) cows had also a higher rate of IMI of 38.2%, 36.7%, and 38.2%, respectively. Conclusion: The IMI and type of bacteria were the principal factors for SCC variation. Besides, mammary quarter location, age, and parity should be taken into consideration during the inter¬pretation of SCC. [J Adv Vet Anim Res 2020; 7(2.000): 314-319]

OBJECTIVE To compare the performance of 5 synthetic peptide–based ELISAs with that of 3 commercially available immunofluorescent assays (IFAs) for serologic diagnosis of anaplasmosis and ehrlichiosis in dogs. SAMPLE A convenience set of 109 serum samples obtained before and at various times after inoculation for 23 dogs that were experimentally infected with Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, Ehrlichia chaffeensis, or Ehrlichia ewingii and 1 uninfected control dog in previous studies. PROCEDURES All serum samples were assessed with 5 synthetic peptide–based ELISAs designed to detect antibodies against A phagocytophilum, A platys, E canis, E chaffeensis, and E ewingii and 3 whole organism–based IFAs designed to detect antibodies against A phagocytophilum, E canis, and E chaffeensis. The species-specific seroreactivity, cross-reactivity with the other tick-borne pathogens (TBPs), and diagnostic sensitivity and specificity were calculated for each assay and compared among assays. RESULTS All serum samples obtained from dogs experimentally infected with a TBP yielded positive results on a serologic assay specific for that pathogen. In general, sensitivity was comparable between ELISAs and IFAs and tended to increase with duration after inoculation. Compared with the IFAs, the corresponding ELISAs were highly specific and rarely cross-reacted with antibodies against other TBPs. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that peptide-based ELISAs had enhanced specificity relative to whole organism–based IFAs for detection of antibodies against Anaplasma and Ehrlichia spp, which should facilitate accurate diagnosis and may help detect dogs coinfected with multiple TBPs.

Today, the use of components obtained from plant extracts is rapidly increasing, especially in the pharmaceutical industry. Eight different plants, which are used as winter tea and are frequently consumed among herbal teas, were selected in the study. The aim of study was to investigate the antimicrobial and antioxidant activities of teas obtained from medicinal and aromatic plants such as Linden, Ginger, Cinnamon, Sage, Daisy, Turmeric, Clove and Rosehip. Five different pathogens (Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis, Escherichia coli and Pseudomonas aeruginosa) were selected from common disease-causing pathogens. A total of 21 combinations were made for each plant. Disc diffusion and Minimum inhibition concentration methods were used to determine antimicrobial activity. DPPH (2,2 Difenil-1Pikrohidrozil) method was used to determine antioxidant activity. The amount of total phenolic and tannins contents contained of herbal teas were also determined using the Folin-Ciocalteu reagent (FCR) method.The highest value among the antimicrobial activities of herbal teas (triple combination) was measured against E. faecalis (25.11 mm). The herbal combination with the highest value measured was found in the ginger+cinnamon+clove group. The highest antioxidant value was measured in this mixture (36.8 mg/mL).Because some plants have more bioavailability, these benefits can be suppressed in a mixture. When determining these mixtures, the consumption will be more beneficial for public health, given the recommendations of researchers and experts.

The primary objective of this study was to explore the variability of Streptococcus uberis (S. uberis) isolates by extracting multilocus sequence typing (MLST) data from whole-genome sequencing. The secondary objective was to determine the distribution of the phenotypic antimicrobial resistance (AMR) and the associated AMR genes as well as the virulence gene profiles among sequence types (STs). Sixty-two isolates were recovered from 16 herds in 3 Canadian Maritime Provinces: New Brunswick (14.5%), Nova Scotia (48.3%), and Prince Edward Island (37.1%). Of these, 9, 30, and 23 were recovered from post-calving, lactational samples, and post-mastitis samples, respectively. These 62 S. uberis isolates belonged to 34 STs; 11 isolates were typed to 9 known STs and 51 isolates were classified as one of 25 new STs. Thirteen isolates were part of major clonal complexes (CCs). Post-mastitis isolates contained 10 unique STs, lactational isolates contained 11 unique STs, and post-calving isolates had 3 STs. Each farm had only 1 isolate that was a unique ST except for STs 233, 851, 855, 857, 864, and 866, which were found in multiple cows per herd on more than one farm. ST851 and ST857 were found in each of the 3 sample types, with ST857 found in cows from all 3 Maritime provinces. These results indicate that S. uberis is a diverse non-clonal pathogen with specific STs residing in clonal clusters, carrying multiple AMR genes and virulence, with a diverse phenotypic AMR.

A total of 23,322 specimens collected between 2014 and 2017, froma total of 2,592 cases, were received in Veterinary Research Institute, Ipoh (VRI) from various states in Malaysia and testedfor common bacterial, viral, and parasitic diseases in pigs. The highest occurrence of isolated bacteria from 771 samples whichtested positive were Salmonella (47.38%) and Escherichia coli (15.68%), followed by Staphylococcus (6.62%), Streptococcus (5.57%), Klebsiella pneumonia (4.88%), Pseudomona (3.38%), Acinetobacter (3.14%), Aeromonas (2.79%), Enterobacter (2.44%), one each of Bacillus and Pasteurella multocida (1.74%), Enterococcus (1.39%) and Corynebacterium (1.05%). 1.74% of each bacteria detected were Moxarella, Aspergillus, Burkholderia andChromobacterium. Positive samples tested by ELISA was Japanese encephalitis virus (JEV) (9.15%), Aujezsky disease virus (ADV)(5.37%), porcine cirvo-virus-2 (PCV2) (5.09%) and porcine reproductive and respiratory syndrome virus (PRRSV) (4.52%). Positive amples tested by the molecular test wasPCV2 (1.62%), PRRSV (1.32%) and classical swine fever virus (CSFV) (0.4%). Serology tests were conducted on 11,305 samplesand reported positive for Brucella suis (15.32%), Brucella abortus (0.62%), Brucella melitensis (0.85%), and melioidosis (0.05%). Parasitology analyses on 99 samples. revealed presence of 10.1% coccidia and 1% each of helminths and Sarcocystis. Within the 4-year period, there were no positive samples for porcine parvovirus (PPV), Nipah virus, swine influenza virus (SIV), and bacteria of Johne’s disease and leptospirosis. Continuous assessment is required to establish a comprehensive baseline data of swine diseases in Malaysia.

OBJECTIVE To determine reference intervals for total nucleated cell count, total protein concentration, pH, RBC count, and percentages of neutrophils, lymphocytes, and large mononuclear cells in synovial fluid samples (SFSs) obtained from the carpal and tarsal joints of healthy swine. ANIMALS 54 healthy commercial finisher pigs that had no evidence of lameness or gross joint swelling. PROCEDURES Each pig was anesthetized, and SFSs were collected from 1 carpal and 1 tarsal joint for fluid analysis, cytologic evaluation, bacterial culture, and PCR analyses for common swine joint pathogens. Each pig was euthanized after SFS collection, and synovial tissue samples were collected for histologic assessment. If necessary, postmortem SFSs were collected. RESULTS Overall, 37 of 50 tarsal and 46 of 53 carpal SFSs met inclusion criteria of sufficient volume, no gross blood contamination, and negative results of bacterial culture and PCR analyses, and were from joints with histologically normal synovial tissues. For the carpal and tarsal joints, upper reference limits were as follows: total nucleated cell count, 3,281 cells/μL and 2,368 cells/μL, respectively; total protein concentration, 3.6 g/dL and 3.6 g/dL, respectively; pH, 7.2 and 7.0, respectively; RBC count, 0.8 × 10(6) cells/μL and 0.1 × 10(6) cells/μL, respectively; and percentage of neutrophils, 46.5% and 33.7%, respectively; percentage of lymphocytes, 40.6% and 56.3%, respectively; and percentage of large mononuclear cells, 92.0% and 95.3%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Results have provided reference intervals for selected variables in SFSs obtained from the carpal and the tarsal joints of healthy swine, which should be useful in diagnostic investigations of swine lameness and arthritis.

The objective of this study was to compare clinical, microbiologic, immunologic, and pathologic parameters in pigs each concurrently administered porcine reproductive and respiratory syndrome virus (PRRSV), Mycoplasma hyopneumoniae, and porcine circovirus type 2 (PCV2) vaccine from 1 of 2 commercial sources at 21 days of age and challenged with field strains of each of the 3 pathogens. Pigs were challenged with PRRSV and M. hyopneumoniae at 42 days of age (-14 days post-challenge, dpc) followed by a challenge with PCV2 at 56 days of age (0 dpc). Significant differences were observed between vaccinated challenged and unvaccinated challenged groups in clinical (average daily gain and clinical signs), microbiologic (viremia and nasal shedding), immunologic (antibodies and interferon-γ secreting cells), and pathologic (lesions) outcomes. Significant differences were observed among the 3 vaccinated challenged groups in microbiologic (nasal shedding of M. hyopneumoniae and viremia of PCV2) and immunologic (M. hyopneumoniae- and PCV2-specific interferon-γ secreting cells) outcomes. The vaccination regimen for PRRSV vaccine, M. hyopneumoniae vaccine, and PCV2 vaccine is efficacious for controlling triple challenge with PRRSV, M. hyopneumoniae, and PCV2 from weaning to finishing period.

There are few reports investigating the characterization of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in dogs in Canada and none from Atlantic Canada. The objectives of this study were to strain type MRSP isolates cultured at a regional diagnostic laboratory using direct repeat unit (dru) typing and to describe their antimicrobial resistance profiles. Ninety-four isolates recovered from dogs between 2010 and 2012 had dru typing, cluster analysis, and antimicrobial susceptibility testing done. The majority of isolates belonged to type dt11a (30.9%), dt10h (24.5%), dt9a (18.1%), and dt11af (10.6%) with the remaining 15.9% of isolates distributed among 13 dru types. The predominant dru types identified were similar in Ontario; however, cluster 9a appears to be less common in Atlantic Canada. A significant difference in the distribution of clusters among Atlantic provinces was detected (P = 0.01). Resistance to ≥ 2 non-β-lactam antimicrobials was observed in 71.4% of the isolates. The MRSP isolates from this study were notably less resistant than those reported in the literature. A more comprehensive study of the MRSP dru types could help further elucidate the distribution of this pathogen in Canada.