Study on supplemental test to improve the detection of bovine tuberculosis in individual animals and herds.

BACKGROUND: Bovine tuberculosis (bTB), is a worldwide disease caused by Mycobacterium bovis (M. bovis). The success of bTB eradication and control programs is based on early detection and the removal of reactors from a herd thus routine testing and cull strategy have been applied globally. Since the late nineteenth century, the Tuberculin Skin Test (TST) has been the primary antemortem test available to support bTB eradication campaigns. Due to the TST limitations in terms of Se and Sp, the credibility of the diagnosis is frequently questioned given the occurrence of false-positive and false-negative reactions, therefore, it is necessary to confirm reactive animals using other methods, ensuring the reliability of the diagnosis. The aim of this study was to evaluate the sensitivity and specificity of a multiplex enzyme-linked immunosorbent assay (ELISA) relative to the tuberculin test used for the diagnosis of tuberculosis in cattle in Brazil. RESULTS: Lack of agreement between comparative cervical tuberculin test and ELISA IDEXX TM was observed. The 2 animals positive on the comparative cervical tuberculin test did not react at the ELISA IDEXX TM and 22 negative reactors by comparative cervical tuberculin test were positive by the ELISA IDEXX TM. The ELISA IDEXX TM showed sensitivity that is significantly lower than the official screening test the single cervical tuberculin. ELISA IDEXX TM also detected infected animals and herds undetected by the comparative cervical tuberculin test. The parallel use of comparative cervical tuberculin test and ELISA IDEXX TM increased sensitivity and the feasibility bTB screening. CONCLUSIONS: The results obtained here suggest that the ELISA IDEXX TM may be a supplemental test for the detection of Mycobacterium bovis infection in regions without routine testing and slaughter, where the disease generally progresses to more advanced stages and antibody responses are likely to be more prevalent. Evidence to support the validation of the ELISA IDEXX™ as a supplemental test for bTB eradication programs was provided.

Prevalence, distribution, and risk factors for canine blastomycosis in Michigan, USA.

Blastomycosis is a systemic fungal infection that most commonly affects dogs and humans. The disease is thought to be endemic in southern regions of Michigan, USA, but epidemiologic investigations have not been reported in detail for this state. The primary aims of this study were to investigate the prevalence and distribution of canine blastomycosis cases in Michigan and to identify risk factors for infection. Over 200 primary care veterinary clinics throughout the state were surveyed regarding blastomycosis prevalence, and demographic information was obtained from medical records of affected dogs that were evaluated at these clinics. A retrospective case control study was conducted for an additional 49 dogs with blastomycosis that were evaluated at specialty referral centers located in the southern mid-Michigan region. Prevalence rates were calculated for each county, and cases were mapped using geocoding software. Univariable and multivariable analyses were used to identify risk factors for infection. Prevalence rates were ≥100 cases per 100,000 dogs in five counties. Most blastomycosis cases originated from the Upper Peninsula or from a high-density area in the northern Lower Peninsula. Multivariable regression analysis identified travel or residence north of the 45th parallel as a strong risk factor for infection (P < .001). Blastomycosis was uncommon in southern counties. These results refute previous speculations and should be of value to both human and animal health. Given that many heightened risk areas are popular tourist destinations, practitioners across the USA should be mindful of the spatial distribution of blastomycosis in Michigan.

Within-Farm Changes in Dairy Farm-Associated Salmonella Subtypes and Comparison to Human Clinical Isolates in Michigan, 2000-2001 and 2009.

Temporal changes in the distribution of Salmonella subtypes in livestock populations may have important impacts on human health. The first objective of this research was to determine the within-farm changes in the population of subtypes of Salmonella on Michigan dairy farms that were sampled longitudinally in 2000-2001 and again in 2009. The second objective was to determine the yearly frequency (2001 through 2012) of reported human illnesses in Michigan associated with the same subtypes. Comparable sampling techniques were used to collect fecal and environmental samples from the same 18 Michigan dairy farms in 2000-2001 and 2009. Serotypes, multilocus sequence types (STs), and pulsed-field gel electrophoresis (PFGE) banding patterns were identified for isolates from 6 farms where >1 Salmonella isolate was recovered in both 2000-2001 and 2009. The distribution of STs was significantly different between time frames (P < 0.05); only two of 31 PFGE patterns were identified in both time frames, and each was recovered from the same farm in each time frame. Previously reported within-farm decreases in the frequency of multidrug-resistant (MDR) Salmonella were due to recovery of MDR subtypes of S. enterica serotypes Senftenberg and Typhimurium in 2000-2001 and genetically distinct, pansusceptible subtypes of the same serotypes in 2009. The annual frequency of human illnesses between 2001 and 2012 with a PFGE pattern matching a bovine strain decreased for patterns recovered from dairy farms in 2000-2001 and increased for patterns recovered in 2009. These data suggest important changes in the population of Salmonella on dairy farms and in the frequency of human illnesses associated with cattle-derived subtypes.

Thirty-minute screening of antibiotic resistance genes in bacterial isolates with minimal sample preparation in static self-dispensing 64 and 384 assay cards.

In a clinical setting, molecular assays such as polymerase chain reaction offer a rapid means to infer or confirm identity and therapeutic decisions. Accordingly, a number of molecular assays targeting identity and antibiotic resistance (AR) genes have been developed; however, these methods can be technically complex and relatively expensive. Herein, we describe a diagnostic concept utilizing isothermal amplification technology with non-purified heat-lysed cells and self-dispensing cards for testing multiple primers in parallel. This proof-of-concept study, performed with Staphylococcus aureus isolates and associated AR genes, was compared with culture-based susceptibility and quantitative PCR (qPCR). Results demonstrate reduced sample processing steps resulting in a turnaround time (starting from bacterial culture to ending in the antibiotic resistance gene profile) in less than 30 min. For antibiotics tested in which an associated AR gene was targeted on the Gene-Z card, 69% (18/26) of culture-based resistance events were positive for related AR genes. A comparison of loop-mediated isothermal amplification (LAMP) and qPCR assays targeting the same antibiotic resistance genes showed a 98.2% agreement in terms of presence and absence calls. Identity-based discrepancies between conventional (phenotypic) and molecular (genotypic) results were further resolved, and we were able to demonstrate higher accuracy in identification with the molecular analysis.

Comparison of PCR versus culture for detection of Mycobacterium bovis after experimental inoculation of various matrices held under environmental conditions for extended periods.

The purpose of this study was to compare the performance of a molecular detection technique (nested PCR) with that of mycobacterial culture in the detection of Mycobacterium bovis DNA in a set of 687 samples of experimentally inoculated environmental substrates (hay, soil, corn, water) exposed to natural weather conditions in Michigan. Four replicates of each substrate were used; half were autoclaved for sterilization, all were inoculated with 50,000 CFU of M. bovis isolated from Michigan livestock, and all were placed in outdoor enclosures, with half under shade and the other half exposed to direct sunlight. Samples were tested for the presence of M. bovis during one 12-month period, with monthly sample testing and during three 12-week periods (winter, spring, summer) with weekly sample testing. Samples were subjected to mycobacterial culture for isolation of M. bovis and a nested PCR with two primer sets targeting IS6110 to detect M. bovis DNA. In 128 samples tested during the 12-month period, M. bovis was not detectable by culture after 2 months but M. bovis DNA was detectable by PCR for at least 7 months. Of the 559 samples tested during the 12-week periods, PCR detected M. bovis DNA for up to 88 days in all of the sample types. There were no significant differences in the detection of M. bovis between shade and sun samples or between sterile and unsterilized samples, regardless of the detection method (PCR or culture). For use in epidemiologic investigations, the PCR assay was more rapid than mycobacterial culture, was not hindered by contaminating organisms, and detected M. bovis DNA in environment samples much longer after initial contamination than mycobacterial culture did.

Herd-level risk factors associated with fecal shedding of Shiga toxin-encoding bacteria on dairy farms in Minnesota, USA.

This study aimed to identify herd-level risk factors associated with fecal shedding of Shiga toxin-encoding bacteria (STB) on dairy cattle farms in Minnesota, USA. After adjustment for farm size, risk factors included: use of total mixed ration (TMR) for lactating dairy cows [odds ratio (OR) = 3.0; 95% confidence interval (CI): 1.8 to 5.1], no use of monensin for weaned calves (OR = 4.8, 95% CI: 2.5, 9.3), and no use of decoquinate for preweaned calves (OR = 2.2, 95% CI: 1.4, 3.6). Fecal shedding of STB was more common in small herds (< 100 cows, OR = 3.6, 95% CI: 2.1, 6.2) than in large herds (≥ 100 cows). Herd management factors related to cattle feeding practices were associated with fecal shedding of STB.

Facteurs de risque au niveau du troupeau associés à l'excrétion fécale des bactéries encodant la toxine de Shiga dans les fermes laitières du Minnesota, États-Unis. Cette étude avait pour but d'identifier les facteurs de risque au niveau du troupeau associés à l'excrétion fécale de bactéries encodant la shiga-toxine dans les fermes de bovins laitiers au Minnesota, États-Unis. Après un ajustement pour la taille de la ferme, les facteurs de risque incluaient : l'utilisation de la ration mixte totale (RMT) pour les vaches laitières en lactation [rapport de cotes (RC) = 3,0; intervalle de confiance (IC) de 95 % : de 1,8 à 5,1], pas d'utilisation de monensin pour les veaux sevrés (RC = 4,8, IC de 95 % : 2,5, 9,3) et pas d'utilisation de décoquinate pour les veaux présevrés (RC = 2,2, IC de 95 % : 1,4, 3,6). L'excrétion fécale de la bactérie encodant la shiga-toxine était plus commune dans les petits troupeaux (< 100 vaches, RC = 3,6, IC de 95 % : 2,1, 6,2) que dans les grands troupeaux (≥ 100 vaches). Des facteurs de gestion du troupeau se rapportant aux pratiques d'alimentation du bétail ont été associés à l'excrétion fécale de la bactérie encodant la shiga-toxine.(Traduit par Isabelle Vallières).

Per- and polyfluoroalkyl substances: using comparative medicine to understand exposure and adverse health outcomes in people and their pets.

One of the important human health benefits of keeping pets may be to serve as an early warning system for indoor childhood exposure to toxic chemicals such as per- and polyfluoroalkyl substances (PFAS). The stain-resistant properties and environmental stability of PFAS make them a preferred choice for protective coatings and lubricants, and they have been used for years in various manufacturing and industrial processes around the world. Although the use of PFAS has arguably improved many commercial products, they have been linked to adverse health outcomes such as developmental delays, liver damage, immune suppression, disruption of endocrine and reproductive systems, and some cancers. The current body of literature suggests that serum PFAS levels in dogs and cats are analogous to their human counterparts and that household pets experience similar changes in blood chemistry markers. The proximity of small children and household pets to PFAS-treated carpets and floors, in addition to their tendency to put things into their mouths, potentially allows pets to serve as sentinels for household PFAS exposure. To assess the suitability of pets as indicators for exposure, researchers need to understand the most likely sources of PFAS exposure for household pets and identify the biomarkers of biological effects in those animals. Understanding these parameters may alert veterinary clinicians to potential sources of contamination in the home and ultimately protect the lives of the children and animals who live there.

Current per- and polyfluoroalkyl substance (PFAS) research points to a growing threat in animals.

Per- and polyfluoroalkyl substances (PFAS) are man-made chemicals that are colorless, odorless, and oil and water repellent. Their widespread use in manufacturing and industrial processes has resulted in environmental contamination found across the world. Exposure to PFAS can lead to a variety of adverse human health outcomes such as increased cholesterol, liver damage, immune suppression, and disruption of endocrine and reproductive systems. Exposure to this family of chemicals is considered a significant public health threat. Though nearly every human and animal around the world has been exposed, most of what is known regarding health effects and toxicological processes of PFAS in animals stems from human epidemiological and laboratory animal studies. Discoveries of PFAS contamination on dairy farms and concerns for companion animals have increased interest in PFAS research related to our veterinary patients. In the limited studies published to date, PFAS has been demonstrated in serum, liver, kidneys, and milk of production animals and has been linked to changes in liver enzymes, cholesterol levels, and thyroid hormones in dogs and cats. This is further addressed in the companion Currents in One Health by Brake et al, AJVR, April 2023. There is a gap in understanding the routes of exposure, absorption of PFAS, and adverse health effects among our veterinary patients. The purpose of this review is to summarize the current literature on PFAS in animals and discuss the implications for our veterinary patients.

Farm-level associations with the shedding of Salmonella and antimicrobial-resistant Salmonella in U.S. dairy cattle.

Salmonella enterica is the leading cause of foodborne-related deaths and hospitalizations within the United States. Infections caused by antimicrobial-resistant (AMR) strains are associated with higher hospital costs and case fatality. The objective for this study was to determine the association of management practices with the recovery of Salmonella and AMR Salmonella on dairy herds. Individual adult cow fecal samples and/or composite fecal samples were collected from 265 dairy herds in 17 states. Samples were cultured for Salmonella, and the MIC was determined for 15 antimicrobials. Herds were classified as Salmonella positive if at least one isolate was recovered, and AMR Salmonella positive if at least one resistant isolate was recovered. Questionnaires regarding management practices were administered to herd operators, and a subset of practices was selected based on subject knowledge and prior research. Data on preventive and therapeutic antimicrobial usage were included in the analysis. Logistic regression models were used to determine which practices were significantly (p<0.05) associated with each herd classification. A total of 124 and 25 herds were classified as Salmonella positive and AMR Salmonella positive, respectively. Variables significantly associated with Salmonella-positive herds included using sprinklers or misters for heat abatement (OR=2.8; CI: 1.6-4.9), feeding anionic salts to cows (OR=1.9; CI: 1.1-3.5), and feeding ionophores to cows (OR=2.1; CI: 1.2-3.7). Herds that used a broadcast/solid spread had lower odds (OR=0.26; CI: 0.11-0.63) of being Salmonella positive. Herds with at least one resistant isolate were more likely to have used composted/dried manure for bedding relative to herds with only susceptible isolates (OR=3.6; CI: 1.2-11.0). These results can be useful to focus additional research aimed at decreasing the prevalence of Salmonella and AMR Salmonella on U.S. dairy herds.

Seromonitoring of brucellosis in goats and sheep slaughtered at an abattoir in Kampala, Uganda.

We conducted a cross-sectional serologic study at Kampala City abattoir in Uganda on 287 small ruminants (221 goats and 66 sheep) to determine the seroprevalence of brucellosis. The samples were tested using a modified rose bengal test (mRBT) and an indirect ELISA (iELISA). Small ruminant Brucella spp. seropositivity was 18 of 287 (6.3%) by mRBT and 19 of 287 (6.6%) by iELISA. The prevalence of brucellosis by mRBT was non-significantly higher in goats (17 of 221; 7.7%) than in sheep (1 of 66, 1.5%; p = 0.069), and also non-significantly higher by the iELISA in goats (18 of 221; 8.1%) than in sheep (1 of 66, 1.5%; p = 0.057). Brucellosis in slaughtered goats and sheep is a public health hazard to abattoir workers and consumers that calls for control and eradication measures at the farm level, given that testing is not carried out routinely at slaughter points.

Efficacy of oral administration of a modified-live Salmonella Dublin vaccine in calves.

OBJECTIVE: To determine the efficacy a modified-live Salmonella Dublin vaccine administered PO in an extralabel manner in the prevention of diseases associated with Salmonella Dublin infection. DESIGN: Randomized clinical trial. ANIMALS: 288 preweaned Holstein dairy calves on a commercial dairy farm. PROCEDURES: Calves were orally administered either 2 mL of a commercially available, modified-live Salmonella Dublin vaccine (n = 140) or a placebo (148) at 3 and 10 days of age. Signs of diarrhea and depression were recorded daily. Weight gain between 3 days of age and time of weaning was measured. Fecal samples from clinically depressed or diarrheic calves and fresh tissues samples from calves that died were submitted for bacterial culture of Salmonella organisms. RESULTS: Salmonella organisms were isolated from samples of 1.4% (2/140) and 3.4% (5/148) of calves receiving the vaccine and placebo, respectively. Additionally, 57.1% (80/140) and 60.1 % (89/148) of the vaccinated and control calves, respectively, had at least 1 day with an abnormal fecal score. Calves receiving the vaccine and placebo were not significantly different in terms of overall morbidity rate, Salmonella-specific morbidity rate, or average daily gain. Adverse reactions related to administration of the vaccine were not seen. The attenuated vaccine strain was not isolated from any fecal or tissue samples. CONCLUSIONS AND CLINICAL RELEVANCE: This method of vaccination was safe in young Holstein calves, although it was not effective in reducing the incidence of disease or improving weight gain on this farm. However, the power of this study was limited by a low incidence of clinical salmonellosis.

Poultry raising systems and highly pathogenic avian influenza outbreaks in Thailand: the situation, associations, and impacts.

Highly pathogenic avian influenza (HPAI), caused by the virus strain H5N1, currently occurs worldwide with the greatest burden in Southeast Asia where the disease was first reported. In Thailand where the disease was first confirmed in January 2004, the virus had been persistent as a major threat to the poultry industry and human health over the past several years. It was generally hypothesized that the main reason for the disease to circulate in Thailand was the existence of traditional backyard chickens and free-range ducks raising systems. Consequently, this study reviewed the structure of poultry raising systems, the recent outbreaks of HPAI H5N1, the disease association to the backyard and free-grazing poultry production, and consequences of the outbreaks in Thailand. Although the major outbreaks in the country had declined, the sustaining disease surveillance and prevention are still strongly recommended.

Genetic Diversity and Potential Paths of Transmission of Mycobacterium bovis in the Amazon: The Discovery of M. bovis Lineage Lb1 Circulating in South America.

Bovine tuberculosis (bTB) has yet to be eradicated in Brazil. Herds of cattle and buffalo are important sources of revenue to people living in the banks of the Amazon River basin. A better understanding of Mycobacterium bovis (M. bovis) populational structure and transmission dynamics affecting these animals can significantly contribute in efforts to improve their sanitary status. Herein, we sequenced the whole genome of 22 M. bovis isolates (15 from buffalo and 7 from cattle) from 10 municipalities in the region of the Lower Amazon River Basin in Brazil and performed phylogenomic analysis and Single Nucleotide Polymorphism (SNP)-based transmission inference to evaluate population structure and transmission networks. Additionally, we compared these genomes to others obtained in unrelated studies in the Marajó Island (n = 15) and worldwide (n = 128) to understand strain diversity in the Amazon and to infer M. bovis lineages. Our results show a higher genomic diversity of M. bovis genomes obtained in the Lower Amazon River region when compared to the Marajó Island, while no significant difference was observed between M. bovis genomes obtained from cattle and buffalo (p ≥ 0.05). This high genetic diversity is reflected by the weak phylogenetic clustering of M. bovis from the Lower Amazon River region based on geographic proximity and in the detection of only two putative transmission clusters in the region. One of these clusters is the first description of inter-species transmission between cattle and buffalo in the Amazon, bringing implications to the bTB control program. Surprisingly, two M. bovis lineages were detected in our dataset, namely Lb1 and Lb3, constituting the first description of Lb1 in South America. Most of the strains of this study (13/22) and all 15 strains of the Marajó Island carried no clonal complex marker, suggesting that the recent lineage classification better describe the diversity of M. bovis in the Amazon.

Comparison of phenotypic and genotypic antimicrobial profiles in Escherichia coli and Salmonella enterica from the same dairy cattle farms.

Transmission of antimicrobial drug resistance from resistant bacteria to non-resistant strains is an important public health issue. In this study, we have examined the possibility of multiple resistance gene transfer between Escherichia coli and Salmonella in the natural setting. Bacteria isolated from calves concurrently shedding E. coli and Salmonella showed similar antimicrobial drug resistance patterns as measured by a broth dilution method. However, microarray analysis of the antibiotic resistance at the gene level revealed several differences in resistance gene profile. Resistance profiles of E. coli isolated from different farms were closer than the profile of E. coli and Salmonella isolated from the same farm. This shows that the chance of multiple resistance gene transfers between these species is unlikely.

Comparison of three immunodiagnostic assays for antemortem detection of Mycobacterium bovis stimulation in domestic cats.

Mycobacterium bovis causes disease in numerous mammalian species including humans, thus making research, surveillance, and control important in the eradication of tuberculosis. Domestic cats are susceptible to multiple mycobacterial species including Mycobacterium bovis; however, their role in the epidemiology of bovine tuberculosis is not fully documented. The current study was an evaluation of the immune response in specific pathogen-free (SPF) cats stimulated with sensitinogen, a heat-killed M. bovis product, using the rapid test, multiple antigen print immunoassay (MAPIA), and bovine-purified protein derivative (bPPD) single skin test. Six cats were inoculated with sensitinogen subcutaneously on days 0 and 24; 2 noninoculated cats and 49 non-SPF cats were controls. Serial serum samples were collected during 135 days and assayed for M. bovis antibodies by rapid test and MAPIA. On day 123, bPPD skin test was performed and read at 48 and 72 hr. The bPPD test at 72 hr had a mean skin thickness of 0.3 mm for stimulated cats and 0.1 mm for controls. Rapid test identified 4 of 6 stimulated cats after bPPD injection. The MAPIA detected antibody against MPB83, 16/83, 16 kDa, and M. bovis culture filtrate (MBCF) antigens. All assays differentiated between stimulated and control cats; however, 7 of 49 non-SPF control cats had a reaction for either antigen MBCF or 16/83. These preliminary studies show potential for antemortem detection of M. bovis among domestic cats. Additional studies to better characterize virulent M. bovis infection in cats would be of value.

The effect of the time interval between exposures on the susceptibility of chickens to superinfection with Marek's disease virus.

Marek's disease virus (MDV) is ubiquitous within commercial poultry flocks because current vaccines do not prevent MDV infection or transmission. In order for newly-evolved MDV strains to become established within a flock, it seems inevitable that any new strain would need to infect and replicate in chickens previously infected with resident MDV strains. This phenomenon is difficult to detect and there is no clear evidence that it is even possible. Four experiments were performed to demonstrate superinfection and evaluate the effect of time between challenges on the effect of superinfection with the use of two pairs of fully virulent MDV strains that could be discriminated by novel technology: 1) JM/102W and rMd5//38CVI, and 2) rMd5 and rMd5//38CVI. Feather follicle epithelium (FFE), spleen, and tumor samples were collected at single or multiple time points from the same bird to determine the frequency and distribution of each virus present following superinfection, with the use of pyrosequencing and immunohistochemistry. Superinfection was observed in 82 of 149 (55%) FFE samples following short-interval challenge (24 hr) compared to only 6 of 121 (5%) samples following long-interval challenge (13 days), indicating a strong influence of challenge interval. In cases where the first inoculated virus was weak or delayed, the second inoculated virus was detected in 42 of 95 (44%) birds. In tumors from dually challenged birds, the second virus was again present much more often following short-interval challenge (68%) compared to long-interval challenge (11%). Virus mixtures in tumors were less common compared to those in FFE samples. Vaccination with turkey herpesvirus had no significant effect on the virus frequency for either virus pair or challenge time interval, suggesting these conclusions may be applicable to vaccinated chickens in the field. These studies demonstrated superinfection for the first time with two fully virulent MDV strains and suggest that short-interval challenge exposure and/or weak initial exposures may be important factors leading to superinfection--a prerequisite for the establishment of a second virus strain in the population. This model system should be useful to elucidate this important phenomenon further.

An outbreak of multidrug-resistant Salmonella enterica serotype Oranienburg in Michigan dairy calves.

The objectives of this study were to report an outbreak of highly drug-resistant Salmonella enterica serotype Oranienburg in dairy calves, and conduct an epidemiological investigation of Oranienburg identified on a dairy herd during a study to determine whether discontinuing feeding medicated milk replacer to preweaned dairy calves resulted in increased antimicrobial susceptibility in enteric bacteria. Calf fecal samples and swabs of calf and maternity pens were collected monthly over 18 months. Samples were streaked onto XLT-4 agar and characteristic colonies were subjected to biochemical tests to confirm Salmonella. Strain relatedness was examined by Xbal and BlnI pulsed-field gel electrophoresis analysis on 62 randomly selected isolates. Antimicrobial susceptibility testing, using automated microbroth dilution, was conducted using a panel containing tetracycline, amikacin, amoxicillin-clavulanic acid, ampicillin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, cefoxitin, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, and trimethoprim-sulfamethoxazole. A total of 190 Salmonella spp. were isolated from 604 calf and 36 pen samples, of which 86% were Oranienburg and 97% were resistant to at least 9 agents. Environmental isolates had lower levels of resistance than fecal isolates. Pulsed-field gel electrophoresis analysis identified three strains: the most common strain was consistently present before the outbreak and at its peak. One strain was exclusively an environmental isolate, with little antimicrobial resistance. Multiresistant isolates with resistance to ciprofloxacin appeared early in the outbreak, and were replaced by multiresistant isolates with resistance to cephalothin. The differences in strains and resistance patterns suggest that the strains of Oranienburg found in fecal isolates may have different origins from environmental isolates.

Prevalence and patterns of antimicrobial resistance in Campylobacter spp isolated from pigs reared under antimicrobial-free and conventional production methods in eight states in the Midwestern United States.

OBJECTIVE: To compare apparent prevalence and patterns of antimicrobial resistance in Campylobacter spp in feces collected from pigs reared with antimicrobial-free versus conventional production methods in 8 states in the Midwestern United States. DESIGN: Cross-sectional study. SAMPLE POPULATION: 95 swine farms that used antimicrobial-free (n = 35) or conventional (60) production methods. PROCEDURES: Fecal samples from 15 pigs/farm were collected. Biochemical and multiplex-PCR analyses were used to identify Campylobacter spp. The minimal inhibitory concentrations of erythromycin, azithromycin, ciprofloxacin, nalidixic acid, gentamicin, and tetracycline for these organisms were determined by use of a commercially available antimicrobial gradient strip. The data were analyzed by use of population-averaged statistical models. RESULTS: Campylobacter spp were isolated from 512 of 1,422 pigs. A subset (n = 464) of the 512 isolates was available for antimicrobial susceptibility testing. The apparent prevalence of Campylobacter spp isolates from pigs on conventional farms (35.8%) and antimicrobial-free farms (36.4%) did not differ significantly. Resistances to azithromycin, erythromycin, and tetracycline were significantly higher on conventional farms (70.0%, 68.3%, and 74.5%, respectively) than antimicrobial-free farms (20.1%, 21.3%, and 48.8%, respectively). Resistances to azithromycin, erythromycin, and tetracycline declined as the number of years that a farm was antimicrobial-free increased. CONCLUSIONS AND CLINICAL RELEVANCE: Production method did not affect the apparent prevalence of Campylobacter spp on swine farms. However, antimicrobial-free farms had a significantly lower prevalence of antimicrobial resistance. Although cessation of antimicrobial drug use will lower resistance over time, investigation of other interventions designed to reduce resistance levels is warranted.

Retrospective analysis of diagnoses and outcomes of 45 cats with micturition disorders presenting as urinary incontinence.

BACKGROUND: In contrast to dogs, the causes and outcomes of urinary incontinence (UI) in cats are largely unknown. OBJECTIVES: To determine the causes, identify comorbid conditions, and assess outcomes of cats with micturition disorders presenting as UI. ANIMALS: Forty-five cats with UI. METHODS: Retrospective study. Medical records of cats presented from January 2006 to December 2017 were searched using 45 keywords related to UI. History, presenting complaint, and physical examination findings were used to confirm a diagnosis. Cases were categorized based on functional and anatomic localizations. RESULTS: Forty-five cats met inclusion criteria. Spinal cord disease was the most common cause of UI (n = 18), followed by urethral (n = 17), bladder (n = 9), and ureteral (n = 1) disorders. Proportions of voiding and storage phase disorders were similar (53% and 47%, respectively). However, voiding-phase disorders were observed more frequently in males and younger-aged cats (P < .03). Urinary tract infection was detected in 11 of 28 (39%) cats. Outcomes were available in 38/45 cases; 16 cats (42%) regained continence, 3 (8%) improved with treatment, and 19 (50%) remained incontinent or were euthanized. Multiple variable logistic regressions indicated that spinal cord disease was significantly more likely to be associated with poor outcomes compared to bladder or urethral disorders (P < .04). CONCLUSIONS AND CLINICAL IMPORTANCE: Urinary incontinence in cats was associated with a variety of congenital and acquired disorders that affected both phases of micturition with similar frequency. Incontinent cats with spinal cord disorders were common and warrant a more guarded prognosis than do cats with bladder or urethral disorders.

Molecular characterization of Mycobacterium bovis infection in cattle and buffalo in Amazon Region, Brazil.

The aim of this study was to characterize Mycobacterium bovis from cattle and buffalo tissue samples, from two Brazilian states, and to analyse their genetic diversity by spoligotyping. Tissue samples from tuberculosis suspect animals, 57 in Amazonas State (12 cattle and 45 buffaloes) and six from Pará State (5 cattle and one buffalo) from slaughterhouses under State Veterinary Inspection, were isolated in culture medium Stonebrink. The positive cultures were confirmed by PCR and analysed by the spoligotyping technique and the patterns (spoligotypes) were identified and compared at the Mycobacterium bovis Spoligotype Database (http://www.mbovis.org/). There was bacterial growth in 44 (69.8%) of the tissues of the 63 animals, of which PCR for region of differentiation 4 identified 35/44 (79.5%) as Mycobacterium bovis. Six different spoligotypes were identified among the 35 Mycobacterium bovis isolates, of which SB0295, SB1869, SB0121 and SB1800 had already been described in Brazil, and SB0822 and SB1608 had not been described. The most frequent spoligotype in this study (SB0822) had already been described in buffaloes in Colombia, a neighbouring country of Amazonas state. The other identified spoligotypes were also described in other South American countries, such as Argentina and Venezuela, and described in the Brazilian states of Rio Grande do Sul, Santa Catarina, São Paulo, Minas Gerais, Mato Grosso do Sul, Mato Grosso and Goiás, indicating an active movement of Mycobacterium bovis strains within Brazil.