RESUMEN

Streptococcus uberis is a common bovine mastitis pathogen in dairy cattle. The rapid identification and characterization of antimicrobial resistance (AMR) in S. uberis plays an important role in its diagnosis, treatment, and prevention. In this study, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to identify S. uberis and screen for potential AMR biomarkers. Streptococcus uberis strains (n = 220) associated with bovine mastitis in northern Thailand were identified using the conventional microbiological methods and compared with the results obtained from MALDI-TOF MS. Streptococcus uberis isolates were also examined for antimicrobial susceptibility using a microdilution method. Principal component analysis (PCA) and the Mann-Whitney U test were used to analyze the MALDI-TOF mass spectrum of S. uberis and determine the difference between antimicrobial-resistant and -susceptible strains. Using MALDI-TOF MS, 73.18% (161/220) of the sampled isolates were identified as S. uberis, which conformed to the identifications obtained using conventional microbiological methods and PCR. Using PCR, antimicrobial-resistant strains could not be distinguished from antimicrobial-susceptible strains for all three antimicrobial agents, i.e., tetracycline, ceftiofur, and erythromycin. The detection of spectral peaks at 7531.20 m/z and 6804.74 m/z was statistically different between tetracycline- and erythromycin-resistant and susceptible strains, respectively. This study demonstrates a proteomic approach for the diagnosis of bovine mastitis and potentially for the surveillance of AMR among bovine mastitis pathogens.

RESUMEN

This study aimed to determine the sensitivity (Se) and specificity (Sp) of a circulating pathogen-specific biomarker (polyketide synthetase 5, Pks5)-based enzyme-linked immunosorbent assay (ELISA) independently or in conjunction with a caudal fold tuberculin (CFT) test for bovine tuberculosis (bTB) screening in dairy cattle. We enrolled 987 dairy cows from 34 herds in Chiang Mai province, Thailand. A conditionally independent Bayesian model with a single population was inferred from the test results. The percentage of positive results for the Pks5-ELISA using 0.4 OD cutoff test and CFT test were 9.0% (89/987) and 10.5% (104/987), respectively. The median of posterior estimates of Se for the Pks5-ELISA test was 90.2% (95% posterior probability interval [PPI] = 76.6-97.4%), while the estimated Sp was slightly higher (median = 92.9, 95% PPI = 91.0-94.5%). The median estimated Se of the CFT test was 85.9% (95% PPI = 72.4-94.6%), while the estimated Sp was higher, with a median of 90.7% (95% PPI = 88.7-92.5%). The posterior estimate for true disease prevalence was 2.4% (95% PPI = 1.2-3.9%). The Pks5-ELISA test yielded characteristics at or above the acceptable standards for bTB detection. Therefore, the pathogen-specific biomarker, Pks5, is a potential detection system for bTB screening and may be applied as an ancillary test together with the currently applied standard method (CFT test) to reinforce the bTB control and eradication programs.

RESUMEN

This study aimed to find methods and interferences and illustrate the pattern of external anal sphincter (EAS) electromyography (EMG) during micturition and to determine reference intervals of electrophysiological bulbocavernosus reflex (EBCR) by using robust statistical methods in healthy spayed female canines. Ten healthy spayed female canines (no breed restriction) with a body weight of 11.3-18 kg were enrolled. EAS EMG during micturition and the EBCR test were performed under light general anesthesia. Altogether 25 out of 34 EAS EMG showed a similar pattern, including low-amplitude high-frequency bursting pattern before voiding, medium- or high-amplitude low-frequency bursting pattern at the beginning of voiding, oscillate medium- and/or high-amplitude low-frequency bursting with a low-amplitude high-frequency bursting pattern during voiding, and high-amplitude high-frequency bursting pattern at the end of voiding. An average of 100 consecutive stimulations of EBCR for one cycle were performed in each dog and another cycle was repeated to ensure reproducibility. The lower and upper limits of the reference interval of EBCR onset latency values and EBCR mean amplitude values were calculated using both standard and robust methods with untransformed and transformed Box-Cox data. The EBCR onset latency was between 13.85 and 27.44 milliseconds, whereas the EBCR mean baseline to peak amplitude was not transformed with Box-Cox transformation. All EBCR compound muscle action potentials started with a negative sharp wave, which tapers from the baseline in the upward direction, showing an upturned bell-shaped curve. In conclusion, this study was possibly the first to examine the method and provide the electrographic pattern of EAS EMG during micturition and reference intervals of EBCR onset latency in spayed female dogs, which may serve as baseline information to help veterinarians differentiate healthy from diseased dogs. Further studies should compare normal dogs and dogs with lower urinary tract abnormalities at different lesion locations.

RESUMEN

Methicillin-resistant Staphylococcus aureus (MRSA) is an opportunistic bacterium that causes many human and animal infections worldwide. MRSA infections are classified as priority infections owing to their high morbidity and mortality, with a significant risk of zoonotic transmission. This study aimed to determine the pooled prevalence of MRSA in dairy cattle farms and its heterogeneity. Relevant studies were retrieved from three databases: PubMed, Web of Science, and Scopus. The pooled prevalence of MRSA in dairy farms was estimated using a random-effects model. Subgroup and meta-regression analyses were used to assess the probable sources of heterogeneity. Sensitivity and publication bias analyses were also performed. A total of 94 articles were eligible for inclusion in this meta-analysis. The pooled prevalence of MRSA was estimated to be 3.81% [95% confidence interval (95% CI) = 2.61-5.20] with significantly high heterogeneity (I 2 = 96.6%, p = 0.00). For the subgroup analysis among continents, the prevalence was highest in Asia (4.89%; 95% CI = 2.88-7.35) and lowest in South America (1.33%, 95% CI = 0.00-5.49). As for the year of publication, MRSA prevalence was highest in reports published from 2015 to 2018 (4.36%, 95% CI = 2.41-6.80) and lowest in reports published before 2015 (2.65%, 95% CI = 0.75-5.52). As for sample type, the prevalence of MRSA in cattle milk (3.91%, 95% CI = 2.64-5.39) was higher than that in other sample types (1.19%, 95% CI = 0.05-3.24). These three factors were not significantly associated with the pooled prevalence of MRSA (p > 0.05). Therefore, the findings of this study indicate that the prevalence of MRSA has been minimal and consistent in dairy cattle farms over time.

RESUMEN

Bovine mastitis could reduce the milk production and the quality of the bulk tank milk (BTM). Antibiotic treatments through intramammary or parenteral methods are being widely used in dairy farms. A cross-sectional study to investigate for general farm management and pre-test the questionnaire was performed in Southwestern Yunnan province, China. A total of 134 dairy farms were included. Milking cows of each farm were determined for the presence of clinical (CM) and sub-clinical (SCM) mastitis using the California Mastitis Test (CMT). Rates of CM and SCM in studied farms ranged from 2-11%, and 24-69%, respectively. The incidence of antibiotic residues in BTM of all farms was very high (32%, 44/134). All antibiotic contaminated samples were from smallholder dairy farms. Factors significantly associated with the presence of antibiotic contamination included farm region, antibiotics usage, persons performing mastitis treatment, and rates of CM. Rates of CM were significantly associated with the farm region, cleanliness of udders before milking, and the number of milking cows. Our results emphasize that the risk factors of dairy farm management should be paid attention, which can reduce mastitis prevalence and antibiotic contamination in BTM in Southwestern China.

RESUMEN

Dairy farming in northern Thailand is expanding, with dairy cattle populations increasing up to 8% per year. In addition, disease outbreaks frequently occur in this region, especially foot-and-mouth disease and bovine tuberculosis. Our goal was to quantify the underlying pattern of dairy cattle movements in the context of infectious disease surveillance and control as movements have been identified as risk factors for several infectious diseases. Movements at district levels within the northern region and between the northern and other regions from 2010 to 2017 were recorded by the Department of Livestock Development. Analyzed data included origin, destination, date and purpose of the movement, type of premise of origin and destination, and type and number of moved cattle. Social network analysis was performed to demonstrate patterns of dairy cattle movement within and between regions. The total numbers of movements and moved animals were 3,906 and 180,305, respectively. Decreasing trends in both the number of cattle moved and the number of movements were observed from 2010 to 2016, with increases in 2017. The majority (98%) of the animals moved were male dairy calves, followed by dairy cows (1.7%). The main purpose of the movements was for slaughter (96.3%). Most movements (67.4%) were shipments from central to northern regions, involving 87.1% of cattle moved. By contrast, 56% of the movements for growing and selling purposes occurred within the northern region, commonly involving dairy cows. Constructed movement networks showed heterogeneity of connections among districts. Of 110 districts, 28 were found to be influential to the movement networks, among which 11 districts showed high centrality measures in multiple networks stratified for movement purposes and regions, including eight districts in the northern and one district in each of the central, eastern, and lower northeastern regions of Thailand. These districts were more highly connected than others in the movement network, which may be important for disease transmission, surveillance, and control.

RESUMEN

This study aimed to estimate the sensitivity (Se) and specificity (Sp) of loop-mediated isothermal amplification (LAMP) and single intradermal tuberculin (SIT) tests for the diagnosis of bovine tuberculosis (bTB) in dairy cattle in Thailand using a Bayesian approach. The SIT test was performed in 203 lactating dairy cattle from nine dairy farms located in Chiang Mai province, Thailand. Milk samples were collected for the LAMP test. Kappa analysis was performed to determine the agreement between the two tests. A one-population conditional independence Bayesian model was applied to estimate the Se and Sp of the two tests. Of 203 dairy cattle, 2 were positive for the SIT test using standard interpretation, whereas 38 were positive for the LAMP test. A poor agreement (kappa = 0) was observed between the two tests. The median Se and Sp of the SIT test using standard interpretation were 63.5% and 99.1%, respectively. The median Se and Sp of the LAMP test were 67.2% and 82.0%, respectively. The estimated true prevalence of bTB was 3.7%. The LAMP test with milk samples can potentially be used as a non-invasive screening test for the diagnosis of bTB in dairy cattle.

RESUMEN

As an alternative to ejaculated semen, epididymal spermatozoa (ES) can be recovered and cryopreserved for use in artificial insemination and other assisted reproductive technologies. However, variabilities in the sperm response to freeze-thaw procedures challenge its inherent value. Therefore, the present study aims to clarify the freezability phenomena in the swamp buffalo ES, where pieces of literature do not abound. Here, we isolated ES from swamp buffaloes using abattoir-derived, post-mortem caudal epididymis by slicing-flushing technique. Following cryopreservation by slow-freezing, ES samples were classified into high (HF) and low freezability (LF) based on their post-thaw total and progressive motilities. Conventional sperm parameters and proteins of interest, such as glutathione S-transferase Mu 3 (GSTM3) and ATP synthase beta subunit (ATP1B1), were assessed and compared between HF and LF. Computer-assisted sperm analysis revealed that nearly all motion and kinematic parameters significantly differed among freezability groups except for wobble, linearity, and straightness. Moreover, intracellular reactive oxygen species production was evident in both HF and LF after fluorescence staining, with the latter having considerably greater malondialdehyde levels than the former. Immunohistochemical labeling demonstrated that both GSTM3 and ATP1B1 proteins were present in the ES and the epididymal tubular epithelium. Although the GSTM3 relative amounts, as analyzed through Western blot, were significantly higher in LF than HF in association with lipid peroxidation, no significant differences were observed in the case of ATP1B1. Such variations in motility, motion and kinematics, oxidative stress status, and specific sperm proteins suggest their potential utility in distinguishing freezability phenotypes in swamp buffalo ES.

Asunto(s)

RESUMEN

This study aimed to (1) investigate the prevalence of bovine tuberculosis (bTB) in slaughtered animals at the Chiang Mai Municipal abattoir in Chiang Mai, Thailand; (2) identify animal-level risk factors for bTB at the abattoir; and (3) evaluate the performance of techniques for bTB detection at the abattoir. From April 2020 to March 2021, 161 animals registered for slaughter were randomly selected for the study. Animal data including age, sex, species, body condition scores, and origins of the animals were collected. Meat inspection was performed by a trained meat inspector. Tissue samples of the lung, liver, and lymph nodes were collected for histopathological diagnosis and polymerase chain reaction (PCR) detection of Mycobacteria and specifically Mycobacterium bovis. The prevalence of bTB during meat inspection and PCR was calculated separately. Animal-level factors affecting bTB were determined using multivariate logistic regression analysis. The performance of meat inspection and PCR was evaluated using a Bayesian approach. The prevalence of bTB was 12.4% (20/161) and 34.8% (56/161) when the disease was diagnosed using meat inspection and PCR, respectively. Buffaloes had a significantly higher risk of being identified as bTB-positive using PCR compared to beef cattle (odds ratio = 2.19; confidence interval = 1.11-4.30). The median of posterior estimates of sensitivity (Se) and specificity (Sp) to detect bTB using meat inspection were 20.8% [95% posterior probability interval (PPI) = 9.1-36.5%] and 87.8% (95% PPI = 79.6-95.4%), respectively. The medians of the posterior estimates of Se and Sp for PCR were 88.6% (95% PPI = 70.5-98.3%) and 94.4% (95% PPI = 84.7-98.8%), respectively. These findings demonstrate that bTB is highly prevalent among slaughtered animals. PCR can be used as an ancillary test for bTB surveillance at abattoirs in Thailand.

RESUMEN

Streptococcus uberis is recognized as an environmental mastitis pathogen in dairy cattle. The varied success rate of antibiotic treatment for S. uberis intramammary infection may be associated with the antimicrobial resistance (AMR) of these bacteria. This observational study aimed to analyze 228 S. uberis strains associated with bovine mastitis in northern Thailand from 2010 to 2017. AMR and AMR genes were determined by the minimum inhibitory concentration (MIC) using a microdilution method and polymerase chain reaction, respectively. The majority of S. uberis strains were resistant to tetracycline (187/228, 82.02%), followed by ceftiofur (44/228, 19.30%), and erythromycin (19/228, 8.33%). The MIC50 and MIC90 of ceftiofur in 2017 were 2-4-fold higher than those in 2010 (P < 0.01). Resistance to tetracycline and ceftiofur significantly increased between 2010 and 2017 (P < 0.05). The most common gene detected in S. uberis was tetM (199/228, 87.28%), followed by ermB (151/228, 66.23 %) and blaZ (15/228, 6.58 %). The association between tetracycline resistance and tetM detection was statistically significant (P < 0.01). The detection rates of tetM significantly increased, while the detection rates of tetO and ermB significantly decreased during 2010-2017. AMR monitoring for bovine mastitis pathogens, especially S. uberis, is necessary to understand the trend of AMR among mastitis pathogens, which can help create an AMR stewardship program for dairy farms in Thailand.

RESUMEN

Staphylococcal food poisoning (SFP), caused by the contamination of staphylococcal enterotoxins, is a common foodborne disease worldwide. The aims of this study were: (1) to investigate classical staphylococcal enterotoxin genes, sea, seb, sec, sed, and see, among Staphylococcus aureus and coagulase-negative staphylococci (CNS) associated with bovine mastitis; (2) to determine the effect of temperature on the expression of classical staphylococcal enterotoxin genes in staphylococci in milk. The detection of classical staphylococcal enterotoxin genes was performed using S. aureus (n = 51) and CNS (n = 47). The expression of classical enterotoxin genes, including sea, seb, sec, and see, was determined during the growth of staphylococci in milk subjected to ultra-high-temperature processing at two different temperatures: 8 °C and room temperature. Classical staphylococcal enterotoxin genes were expressed more frequently in S. aureus (35.30%) than in CNS (12.77%). The sec gene was most frequently detected in S. aureus (29.41%) and CNS (6.38%). Moreover, the expression of sea and sec was significantly higher at room temperature than at 8 °C after 16 h of incubation (p < 0.05). These results emphasize the importance of maintaining the storage temperature of milk below 8 °C to reduce the risk of SFP.

RESUMEN

Methicillin-resistant Staphylococcus aureus (MRSA) from dairy animals could pose a public health concern in the population. The study was designed to determine the prevalence of S. aureus and MRSA associated with mastitis among water buffaloes in the central part of Luzon island, the Philippines, and to investigate its associated factors. Three hundred and eighty-four water buffaloes were examined for mastitis using California mastitis test (CMT). Composite milk samples (n = 93) were collected from buffaloes showing positive reaction with CMT. S. aureus was identified from milk samples using biochemical tests. Cefoxitin disk diffusion assay and PCR detecting mecA gene were performed to identify MRSA isolates. Disk diffusion assay was used to investigate the antimicrobial resistance against 9 antibiotics. The prevalence of S. aureus was 41.94% (39/93). MRSA isolates resistant to cefoxitin were at 25.81% (24/93) but only 37.5% (9/24) harbored the mecA gene. All 24 MRSA isolates were resistant to penicillin while the majority were susceptible to clindamycin, trimethoprim-sulfamethoxazole, gentamycin, tetracycline, rifampicin, ciprofloxacin and chloramphenicol with intermediate susceptibility to erythromycin. Furthermore, 37.5% of the isolates were found resistant to two or more antibiotics. Animal-level factor associated with MRSA infection was the history of mastitis (OR = 3.18, CI = 1.03-9.79, p = 0.040). Herd-level factors associated with the detection of MRSA in milk included herd size (OR = 4.24, CI = 1.05-17.07, p = 0.042) and the presence of other animals (OR = 0.15, CI = 0.04-0.58, p = 0.006). High prevalence of intramammary infection with S. aureus and MRSA in dairy buffaloes was observed in the region. This finding raises the concern of preventing zoonotic spread of MRSA.

RESUMEN

OBJECTIVE: Mastitis is considered as an economically important disease of dairy buffaloes in Asia. This study examined the mastitis milk and nasal swab samples for the detection and genotyping of methicillin-resistant Staphylococcus aureus (MRSA) in water buffaloes. MATERIALS AND METHODS: Staphylococcus aureus was identified based on biochemical tests and Polymerase Chain Reaction (PCR) detection of nuc gene, whereas MRSA on mecA gene. The disc diffusion test was used to determine the antibiotic resistance and staphylococcal cassette chromosome mec (SCCmec), spa, and multilocus sequence typing for the genotyping of isolates. RESULTS: Staphylococcus aureus was detected on 39/93 milk (41.94%) and 27/384 nasal swab (7.03%) samples. However, only nine isolates (23.08%) harbored the mecA gene from milk samples and three isolates (11.11%) from the nasal carriage. All MRSA isolates exhibited resistance to cefoxitin and penicillin, whereas 50% were found resistant to clindamycin. All these isolates were found susceptible to sulfa-trimethoprim and chloramphenicol, whereas the majority of the isolates were susceptible to gentamicin, ciprofloxacin, tetracycline, and rifampicin. The SCCmec types of the MRSA isolates were type IVc (50.00%), type II (8.33%), type I (8.33%), and non-typeable (33.33%). The spa types and sequence type (ST) identified were t019 (ST30), t701 (ST1649), t311 (ST5), t657 (ST1148), t015 (ST508), t1939 (ST12), t800 (ST9), t091 (ST2454), t138 (ST5991), and t1642 (ST5992). CONCLUSION: Milk and nasal swab samples from dairy water buffaloes were found positive for MRSA. The MRSA isolates were still susceptible to most antibiotics tested. Moreover, the genotypes of some MRSA isolates were found similar to some human MRSA strains, suggesting a possible human to animal transmission.

RESUMEN

Bovine mastitis is a major health problem that affects dairy cows and has a negative impact on milk production. The presence of microRNAs in biofluids, such as blood and milk, could play a pivotal role in the detection of bovine mastitis. The purpose of the current study was to determine the levels of microRNA gene expression in milk, in combination with other reported mastitis indicators, as a biomarker of bovine mastitis. Milk samples (n = 171) were obtained from 113 dairy cows with known disease status (i.e., healthy; n = 23 cows, subclinical mastitis; n = 45 cows, or clinical mastitis; n = 45 cows) and analyzed for the presence of MIR24-2, MIR29B-2, MIR146A, MIR148A, MIR155, MIR181A1, MIR184, and MIR223 expression using the real-time PCR (qPCR) method. The expression data were then utilized in the creation of receiver operator characteristic curves (ROC) and further analyzed by the machine learning (ML) methods. MIR29B-2, MIR146A, MIR148A, and MIR155 expression levels differed significantly among the three groups. These potential microRNA biomarkers of mastitis exhibited high sensitivity and specificity. Next, we applied ML algorithm, specifically, a decision tree (DT) model to predict the status of milk based on MIR29B-2 and MIR146A expression levels. The results suggested that MIR29B-2, when used in combination with the California mastitis test (CMT) and days in milk (DIM) data, was applicable for screening and classification of milk samples from cows as healthy, subclinical mastitis, or mastitis. MIR29B-2 appears to have sufficient discriminatory power to enable it to be utilized as a biomarker in cases where the status of a milk sample cannot be determined based on CMT results.

Asunto(s)

RESUMEN

Streptococcus spp. are major pathogenic bacteria associated with massive mortality in tilapia. This study investigated the phenotypic and genotypic characterization of Streptococcus agalactiae (GBS) and Streptococcus iniae (S. iniae) isolated from tilapia in river-based floating cage and earthen pond farms in northern Thailand. Isolates were identified by biochemical and molecular analyses. Capsular typing, enterobacterial repetitive intergenic consensus polymerase chain reaction and multilocus sequence typing were performed to investigate the genetic relatedness. Six and one isolates were confirmed as GBS and S. iniae, respectively. All Streptococcus spp. isolates were obtained from 4 river-based cage farms (4/33), while samples collected from earthen pond farms (N = 28) were negative for streptococcosis. All GBS with serotype Ⅲ and sequence type (ST) 283 was observed. The ß-haemolytic GBS isolates were resistant to five antimicrobials, while the S. iniae was susceptible to all antimicrobials. This study indicates both GBS and S. iniae are the major bacterial pathogens responsible for streptococcosis infection in farmed tilapia of northern Thailand with GBS as dominant species. This survey highlights that the river-based cage farms seriously impact on the healthy development of the tilapia industry.

Asunto(s)

RESUMEN

Streptococcus agalactiae is well recognized to cause a variety of infections in many animal species and humans. We aimed to investigate genetic relatedness of S. agalactiae strains isolated from humans and animal origins, including cattle and fish, using capsular gene typing, pulsed-field gel electrophoresis (PFGE), and multilocus sequence typing techniques. Our results revealed that S. agalactiae strains with capsular type Ia and ST103 were observed from all bovine isolates (17/17) and one human isolate (1/5). S. agalactiae strains with capsular type III and ST283 were detected among isolates from fish (5/5) and from humans (2/5). Two PFGE clusters containing isolates from mixed origins were demonstrated: one cluster of five fish and one human isolate, and another cluster of one bovine and one human isolate. In conclusion, the close genetic relationship among S. agalactiae strains isolated from humans and animal origins was evident.

Asunto(s)

RESUMEN

BACKGROUND: The objective of this study was to determine the sensitivity (Se) and specificity (Sp) of bovine tuberculosis (bTB) screening tests including a single intradermal tuberculin (SIT) test, interferon gamma (IFN-γ) assay, and a commercial ELISA test (M. bovis Ab) in dairy cattle, under field conditions, using a Bayesian approach. RESULTS: The study population consisted of 128 dairy cows from 25 bTB-infected herds in Chiang Mai and Chiang Rai provinces, Thailand. A single-population Bayesian model was implemented assuming conditional dependence between the SIT test and IFN-γ assays. The 95% posterior probability interval (PPI) of the SIT test (severe interpretation) Se ranged from 75.3 to 95.2% (median = 87.6%), while the Sp was slightly lower (median = 83.6%, PPI = 74.2-92.8%). The IFN-γ assay Se was moderate and the 95% PPI ranged from 38.6 to 74.4% (median = 55.7%) with higher Sp (median = 93.5.4%, PPI = 87.0-98.1%). The M. bovis Ab ELISA Se was low, with 95% PPI ranging between 30.0 and 71.2% (median = 47.4%); however, the Sp was high (median = 90.9%, PPI = 84.5-95.5%). CONCLUSION: The SIT test sensitivity was similar to that demonstrated in other regions and can, therefore, be used effectively as part of control programs in this area. The IFN-γ and M. bovis Ab ELISA assays can be applied as supplementary techniques. The test performance of these tests when used as single tests without confirmation, however, are expected to continue to challenge disease eradication efforts.

Asunto(s)

RESUMEN

AIM: To investigate gene expression of microRNA (miRNA) milieus (MIRLET7E, MIR17, MIR24-2, MIR146A, and MIR181C), inflammatory cytokine genes (interleukin 1ß [IL1B], IL6, CXCL8, and tumor necrosis factor [TNF]), and the pathogen receptor toll-like receptor (TLR4) in bovine neutrophils under quercetin supplementation. MATERIALS AND METHODS: Isolated bovine neutrophils were incubated with bacterial lipopolysaccharide under quercetin treatment or left untreated. Real-time polymerase chain reaction was performed to determine the expression of the miRNAs and messenger RNA (mRNA) transcripts in neutrophils. RESULTS: Quercetin-treated neutrophils exhibited a remarkable suppression in MIR24-2, MIR146A, and MIR181C expression. Similarly, mRNA expression of IL1B, IL6, CXCL8, TLR4, and TNF genes noticeably declined in the quercetin group. Many proinflammatory genes (IL1B, IL6, and CXCL8) and the pathogen receptor TLR4 had a negative correlation with MIR146A and MIR181C as revealed by Pearson correlation. CONCLUSIONS: Interaction between cognate mRNAs and miRNAs under quercetin supplementation can be summarized as a positive or negative correlation. This finding may help understand the effects of quercetin either on miRNA or gene expression during inflammation, especially as a potentially applicable indicator in bovine mastitis.

RESUMEN

The objective of this case-control study was to identify farm-level risk factors associated with bovine tuberculosis (bTB) in dairy cows in northern Thailand. Spatial analysis was performed to identify geographical clustering of case-farms located in Chiang Mai and Chiang Rai provinces in northern Thailand. To identify management factors affecting bTB status, a matched case-control study was conducted with 20 case-farms and 38 control-farms. Case-farms were dairy farms with at least single intradermal tuberculin test- (SIT-) reactor(s) in the farms during 2011 to 2015. Control-farms were dairy farms with no SIT-reactors in the same period and located within 5 km from case-farms. Questionnaires were administered for data collection with questions based on epidemiological plausibility and characteristics of the local livestock industry. Data were analyzed using multiple logistic regressions. A significant geographic cluster was identified only in Chiang Mai province (p < 0.05). The risk factor associated with presence of SIT-reactors in dairy herds located in this region was purchasing dairy cows from dealers (OR = 5.85, 95% CI = 1.66-20.58, and p = 0.006). From this study, it was concluded that geographic clustering was identified for dairy farms with SIT-reactors in these provinces, and the cattle movements through cattle dealers increased the risks for SIT-reactor farm status.

RESUMEN

Microbiological culture of milk samples has been used as a standard diagnosis for Mycoplasma mastitis. This technique is effective in isolating mollicutes that are Mycoplasma-like; however, isolates may be misinterpreted as Acholeplasma species, which are indistinguishable from Mycoplasma species by culture. A study to contrast the abilities of 2 culture-based tests, digitonin and nisin disc diffusion assays and a conventional polymerase chain reaction (PCR) technique, to discriminate between Mycoplasma and Acholeplasma was performed using 16S ribosomal RNA gene partial sequencing as the gold standard of comparison. A total of 288 bovine mollicute field isolates (248 from milk and 40 from other organ sites) and 13 reference strains were tested. Results obtained from the digitonin disc diffusion assay when it was performed with all field isolates were 92.7% and 99.0% in agreement with the gold standard using 5 mm and 3 mm of zone of growth inhibition as thresholds, respectively. Considering only milk isolates, agreements between the digitonin disc diffusion assay with the gold standard were 97.2% and 100% using 5 mm and 3 mm of zone of growth inhibition as thresholds, respectively. Culture identification using the nisin disc diffusion assay and the PCR was in a 100% agreement with the gold standard. Comparable results using culture-based nisin and digitonin disc diffusion assays, and PCR, to distinguish Mycoplasma and Acholeplasma species was found, especially for isolates from bovine milk.

Asunto(s)