RETROSPECTIVE REVIEW OF COPPER SULFATE IMMERSION TREATMENT IN MARINE TELEOSTS DURING QUARANTINE AT THE NATIONAL AQUARIUM OF BALTIMORE FROM 2004 TO 2016.

Copper sulfate immersion is common for the prevention and treatment of Cryptocaryon irritans during quarantine of marine teleosts. The National Aquarium in Baltimore has followed a consistent copper sulfate protocol for marine teleost quarantine since 2004. The protocol used copper sulfate pentahydrate as a slow drip to increase copper ions over 3-5 days to a level of 0.18-0.21 mg/L. This level was maintained for 21 days, and then copper ions were rapidly removed with activated carbon filtration and water changes. Quarantine records from 2004-2016 were used to examine mortality of marine teleosts during copper treatment and identify factors that might have influenced mortality. The following records were excluded: brackish and freshwater teleosts (salinity <25 g/L); long-term treatment at subtherapeutic levels (<0.18 mg/L); intentional short courses (<14 days); and use outside of quarantine. Species, system volume, temperature, parasitic outbreaks, concurrent medications, and water quality concerns were evaluated. During this period, 4,835 individual teleosts belonging to 347 different species were treated. From 2004 to 2016, mortality during copper treatment was 4.1% (199/4,835 individuals) and was higher when treatment was started during the first week of quarantine (7.7%, 68/884) rather than later (3.3%, 131/3,951 individuals). Of the mortalities, 24.1% (48/199) occurred during the initial subtherapeutic period, and 75.9% (151/199) occurred during the therapeutic period. No mortalities occurred in 75.5% (262/347) of species during copper treatment. When using a similar methodology, copper sulfate is a safe immersion for quarantine of marine teleosts. Mortalities during copper treatment can be reduced by increasing copper ion levels to therapeutic ranges more slowly (e.g., over 7 days) and starting copper treatment after the first week of quarantine.

Cognitive Function Among Antiretroviral Treatment-Naive Individuals Infected With Human Immunodeficiency Virus Type 1 Subtype G Versus CRF02_AG in Nigeria.

Background: Human immunodeficiency virus type 1 (HIV-1) subtype has been shown to be associated with disease progression. We compared cognitive function between individuals infected with HIV-1 subtype G and CRF02_AG in Nigeria. Methods: For this cross-sectional study, samples were analyzed from 146 antiretroviral-naive participants. Genotypic analysis of plasma HIV RNA was performed by nested polymerase chain reaction of protease and reverse transcriptase genes, and sequences were aligned with curated HIV-1 subtype references. Cognitive status was determined using demographically adjusted T scores and global deficit score (GDS) obtained from a comprehensive neuropsychological test battery. Results: A total of 76 (52.1%) participants were infected with CRF02_AG, 48 (32.8%) with subtype G, and 22 (15.1%) with other HIV-1 strains. In a multivariable linear regression adjusting for plasma HIV RNA, CD4 count, and depression score, mean global T score was lower among subtype G-infected compared with CRF02_AG-infected participants (mean difference, -3.0 [95% confidence interval {CI}, -5.2, to -.7]; P = .011). Also, T scores were significantly lower among subtype G- than CRF02_AG-infected participants for the speed of information processing, executive function, and verbal fluency ability domains. Adjusting for similar variables in a logistic regression, the odds of global cognitive impairment (GDS ≥0.5) were 2.2 times higher among subtype G compared with CRF02_AG-infected participants (odds ratio, 2.2 [95% CI, .9-5.4]; P = .078). Conclusions: Cognitive performance was significantly worse among antiretroviral-naive individuals with HIV-1 subtype G vs CRF02_AG infection. Further studies are required to characterize the mechanistic basis for these differences.

Plasma HIV RNA level is associated with neurocognitive function among HIV-1-infected patients in Nigeria.

Plasma HIV RNA level has been shown to correlate with HIV disease progression, morbidity, and mortality. We examined the association between levels of plasma HIV RNA and cognitive function among patients in Nigeria. A total of 179 HIV-1-infected participants with available plasma HIV RNA results and followed longitudinally for up to 2 years were included in this study. Blood samples from participants were used for the measurement of plasma HIV RNA and CD4+ T cell count. Utilizing demographic and practice effect-adjusted T scores obtained from a seven-domain neuropsychological test battery, cognitive status was determined by the global deficit score (GDS) approach, with a GDS ≥ 0.5 indicating cognitive impairment. In a longitudinal multivariable linear regression analysis, adjusting for CD4 cell count, Beck's Depression Score, age, gender, years of education, and antiretroviral treatment status, global T scores decreased by 0.35 per log10 increase in plasma HIV RNA [p = 0.033]. Adjusting for the same variables in a multivariable logistic regression, the odds of neurocognitive impairment were 28% higher per log10 increase in plasma HIV RNA (OR 1.28 [95% CI 1.08, 1.51]; p = 0.005). There were statistically significant associations for the speed of information processing, executive, and verbal fluency domains in both linear and logistic regression analyses. We found a significant association between plasma HIV RNA levels and cognitive function in both baseline (cross-sectional) and longitudinal analyses. However, the latter was significantly attenuated due to weak association among antiretroviral-treated individuals.

Peripheral blood lymphocyte HIV DNA levels correlate with HIV associated neurocognitive disorders in Nigeria.

Mononuclear cells play key roles in the pathogenic mechanisms leading to HIV-associated neurocognitive disorders (HANDs). We examined the association between HIV DNA within peripheral blood mononuclear cell (PBMC) subsets and HAND in Nigeria. PBMCs were collected at baseline from 36 antiretroviral naive participants. CD14+ cells and T&B lymphocyte fractions were isolated by, respectively, positive and negative magnetic bead separation. Total HIV DNA within CD14+ and T&B cells were separately quantified using real-time PCR assay targeting HIV LTR-gag and cell input numbers determined by CCR5 copies/sample. Utilizing demographically adjusted T scores obtained from a 7-domain neuropsychological test battery, cognitive status was determined by the global deficit score (GDS) approach, with a GDS of ≥0.5 indicating cognitive impairment. In a linear regression adjusting for plasma HIV RNA, CD4 and lymphocyte count, Beck's depression score, and years of education, there was 0.04 lower log10 HIV DNA copies within T&B lymphocytes per unit increase in global T score (p = 0.02). Adjusting for the same variables in a logistic regression, the odds of cognitive impairment were 6.2 times greater per log10 increase in HIV DNA within T&B lymphocytes (p = 0.048). The association between cognitive impairment and HIV DNA within CD14+ monocytes did not reach statistical significance. In this pretreatment cohort with mild cognitive dysfunction, we found a strong association between levels of HIV DNA within the lymphocyte subset and HAND independent of plasma HIV RNA. These findings likely reflect the neurologic impact of a larger HIV reservoir and active viral replication.

Microbiota that affect risk for shigellosis in children in low-income countries.

Pathogens in the gastrointestinal tract exist within a vast population of microbes. We examined associations between pathogens and composition of gut microbiota as they relate to Shigella spp./enteroinvasive Escherichia coli infection. We analyzed 3,035 stool specimens (1,735 nondiarrheal and 1,300 moderate-to-severe diarrheal) from the Global Enteric Multicenter Study for 9 enteropathogens. Diarrheal specimens had a higher number of enteropathogens (diarrheal mean 1.4, nondiarrheal mean 0.95; p<0.0001). Rotavirus showed a negative association with Shigella spp. in cases of diarrhea (odds ratio 0.31, 95% CI 0.17-0.55) and had a large combined effect on moderate-to-severe diarrhea (odds ratio 29, 95% CI 3.8-220). In 4 Lactobacillus taxa identified by 16S rRNA gene sequencing, the association between pathogen and disease was decreased, which is consistent with the possibility that Lactobacillus spp. are protective against Shigella spp.-induced diarrhea. Bacterial diversity of gut microbiota was associated with diarrhea status, not high levels of the Shigella spp. ipaH gene.

Cost-effectiveness of point-of-care digital chest-x-ray in HIV patients with pulmonary mycobacterial infections in Nigeria.

BACKGROUND: Chest-x-ray is routinely used in the diagnosis of smear negative tuberculosis (TB). This study assesses the incremental cost per true positive test of a point-of-care digital chest-x-ray, in the diagnosis of pulmonary mycobacterial infections among HIV patients with presumed tuberculosis undetected by smear microscopy. METHODS: Consecutive patients with clinical suspicion of pulmonary tuberculosis were serially tested for Human immunodeficiency virus (HIV), their sputum examined for Acid Fast Bacilli then cultured in broth and solid media. Cultures characterized as tuberculous (M.tb) and non-tuberculous (NTM) mycobacteria by Hain assays were used as gold standards. A chest-x-ray was classified as: (1) consistent for TB, (2) not consistent for TB and (3) no pathology. RESULTS: Of the 1391 suspected cases enrolled, complete data were available for 952 (68%): 753/952 (79%) had negative smear tests while 150/753 (20%) had cultures positive for TB. Of those, 82/150 (55%) had chest-x-ray signs consistent with TB and 29/82 (35%) were positive for HIV. Within the co-infected, 9/29 (31%) had NTM infections. Among all suspects, the cost per positive case detected using smear microscopy test was $52.84; the overall incremental cost per positive case using chest-x-ray in smear negatives was $23.42, and in smear negative, HIV positive patients the cost was $15.77. CONCLUSION: Point-of-care chest-x-ray is a cost-effective diagnostic tool for smear negative HIV positive patients with pulmonary mycobacterial infection.

Nonmedical factors associated with feather picking in pet psittacine birds.

A nested case-control study was performed to determine nonmedical risk factors associated with feather picking in psittacine birds. Forty-two case birds, reported by their owners to pick their feathers, and 126 unaffected birds were compared. The odds of feather picking were higher in 2 species categories, African grey parrots (Psitticus erithacus, adjusted odds ratio [ORadj = 8.4, P < .001) and cockatoos (Cacatua species, ORadj = 12.7, P < .001). The odds of feather picking also were higher for birds that were out of their cages more than 8 hours per day (ORadj = 7.4, P < .001) and for birds that had been taken in by the owner as a "rescue" (ORadj = 4.7, P < .01). The odds of feather picking decreased by almost 90% (ORadj = 0.1, P < .005) for birds that interacted with people at least 4 hours a day. These findings identify characteristics that practitioners may want to include when asking bird owners about behavioral history and may be useful in focusing future research regarding this behavior.

Determining diagnostic sensitivity loss limits for sample pooling in duplex rtPCR surveillance testing: Theileria orientalis and Anaplasma marginale.

To expand surveillance testing capacity through sample pooling, a thorough understanding is needed of how sample dilution through pooling affects the sensitivity of candidate assays. We validated a robust and representative framework for assessing the dilution effect of sample pooling using duplex rtPCR surveillance of Theileria orientalis and Anaplasma marginale, both of which are causative agents of severe anemia in cattle and a serious threat to the cattle industry in Virginia and many other states. We used 200 known-positive samples with Ct values representative of typical surveillance results in a series of pools in which we re-tested each sample individually, followed by each sample diluted in equal volumes with negative samples to make pools of 2, 4, 6, 8, and 10 total samples. We compared the Ct values of the individual positives with the Ct values of each pool size to determine if Ct values increase past the limit of detection in the 45-cycle assay. We observed a maximum of 2% sensitivity loss (no more than 2 of 100 samples returned a false-negative result) for both T. orientalis and A. marginale during the pooling series, with lower-than-expected average Ct increase and sensitivity loss. We conclude that pooling up to 10 samples would be acceptable for regional surveillance of T. orientalis and A. marginale using our rtPCR assay. The described strategy is applicable to validate pooling for a wide range of single and duplex rtPCR assays, which could expand efficient disease surveillance.

Theileria orientalis Ikeda infection does not negatively impact growth performance or breeding soundness exam results in young beef bulls at bull test stations.

Introduction: Theileria orientalis Ikeda genotype is an emerging cattle disease in the US. Since 2017, when T. orientalis Ikeda was discovered in beef cattle in two counties in Virginia, cattle infections have risen to include ~67% of Virginia counties and 14 states. Consistent with New Zealand studies, many infected herds in Virginia were >90% positive upon initial testing without overt evidence of infection. Central bull tests present a unique opportunity to study the effects of T. orientalis Ikeda infections, as bulls from multiple source herds are consolidated. The objective of this study was to determine if infection with T. orientalis Ikeda affected the average daily gain (ADG), adjusted yearling weight (AYW) and breeding soundness of bulls at two test stations in Virginia over a period of years. Materials and methods: The bulls were fed and housed similarly to compare their growth performance and breeding soundness. For T. orientalis Ikeda testing, DNA was extracted from whole blood for quantitative polymerase chain reaction. Results: The number of bulls infected with T. orientalis Ikeda at initial delivery to the stations increased significantly over the years studied. Multivariable linear regression models, using Angus bulls from Virginia test stations, indicated no significant effect on ADG or AYW in bulls that became test positive during the test or were positive for the duration, compared to Angus bulls that were negative for the duration. At LOC A, the odds of passing a breeding soundness exam (BSE) were not significantly different for bulls that turned positive during the test or were positive for the duration, compared to bulls that were negative for the duration of the test. At LOC B, bulls that became positive during the test were 2.4 times more likely (95% CI: 1.165-4.995, p = 0.016) to pass their BSE compared to bulls that remained negative throughout the test. Discussion: We do not suppose that an obscured infection of T. orientalis Ikeda is protective for bulls to pass a BSE. However, this study demonstrates an obscured infection of T. orientalis Ikeda does not negatively affect weight gain or achievement of a satisfactory BSE rating at the central bull test stations in Virginia.

Development of a case definition for clinical feline herpesvirus infection in cheetahs (Acinonyx jubatus) housed in zoos.

The identification of feline herpesvirus (FHV) infected cheetahs (Acinonyx jubatus) and characterization of shedding episodes is difficult due to nonspecific clinical signs and limitations of diagnostic tests. The goals of this study were to develop a case definition for clinical FHV and describe the distribution of signs. Medical records from six different zoologic institutions were reviewed to identify cheetahs with diagnostic test results confirming FHV. Published literature, expert opinion, and results of a multiple correspondence analysis (MCA) were used to develop a clinical case definition based on 69 episodes in FHV laboratory confirmed (LC) cheetahs. Four groups of signs were identified in the MCA: general ocular signs, serious ocular lesions, respiratory disease, and cutaneous lesions. Ocular disease occurred with respiratory signs alone, with skin lesions alone, and with both respiratory signs and skin lesions. Groups that did not occur together were respiratory signs and skin lesions. The resulting case definition included 1) LC cheetahs; and 2) clinically compatible (CC) cheetahs that exhibited a minimum of 7 day's duration of the clinical sign groupings identified in the MCA or the presence of corneal ulcers or keratitis that occurred alone or in concert with other ocular signs and skin lesions. Exclusion criteria were applied. Application of the case definition to the study population identified an additional 78 clinical episodes, which represented 58 CC cheetahs. In total, 28.8% (93/322) of the population was identified as LC or CC. The distribution of identified clinical signs was similar across LC and CC cheetahs. Corneal ulcers and/or keratitis, and skin lesions were more frequently reported in severe episodes; in mild episodes, there were significantly more cheetahs with ocular-only or respiratory-only disease. Our results provide a better understanding of the clinical presentation of FHV, while presenting a standardized case definition that can both contribute to earlier diagnoses and be used for population-level studies.

Career stage differences in mental health symptom burden and help seeking among veterinarians during COVID-19.

OBJECTIVE: To explore veterinarians' mental health symptom burden during COVID-19 and identify differences in symptom burden, social support, help seeking, and incentives and barriers associated with receiving help across career stages. SAMPLE: Online survey responses from 266 veterinarians between June 4 and September 8, 2021. PROCEDURES: Respondents were grouped by career stage (early [< 5 years of experience], middle [5 to 19 years of experience], or late [≥ 20 years of experience]), and results were compared across groups. RESULTS: Of the 262 respondents who reported years of experience, 26 (9.9%) were early career, 130 (49.6%) were midcareer, and 106 (40.4%) were late career. The overall mean anxiety and depression symptom burden score was 3.85 ± 3.47 (0 to 2 = normal; 3 to 5 = mild; 6 to 8 = moderate; and 9 to 12 = severe), with 62 of 220 (28.1%) respondents reporting moderate to severe symptom burden. Most (164/206 [79.6%]) reported not accessing behavioral health providers, and of these, 53.6% (88/164) reported at least mild symptom burden. There were significant differences in both symptom burden and mental health help-seeking intentions across career stages, with early- and midcareer (vs late-career) veterinarians reporting higher symptom burden (P = .002) and midcareer (vs late-career) veterinarians reporting higher help-seeking intentions (P = .006). Barriers and incentives for seeking mental health care were identified. CLINICAL RELEVANCE: Findings revealed differences in symptom burden and intentions to seek mental health care across veterinary career stages. Incentives and barriers identified serve to explain these career stage differences.

Veterinarians show resilience during COVID-19: challenges faced and successful coping strategies.

OBJECTIVE: To identify challenges veterinarians faced during the COVID-19 pandemic, explore how they coped, identify coping strategies associated with greater resilience, and determine incentives and barriers to performing healthy coping behaviors. SAMPLES: 266 surveys completed by veterinarians in the Potomac region. PROCEDURES: A cross-sectional survey was distributed electronically through veterinary medical boards and professional associations between June and September 2021. RESULTS: Most survey responses came from veterinarians working in Maryland (128/266 [48%]) and Virginia (63/266 [24%]) who were predominantly white (186/266 [70%]), female (162/266 [61%]), and working in small-animal clinical practice (185/266 [70%]). The greatest workplace challenges experienced were increased workloads (195/266 [73%]) and reevaluating existing workflows (189/266 [71%]). Separation from loved ones (161/266 [61%]) was the greatest personal challenge. Of the veterinarians who completed the 10-point Connor-Davidson Resilience Scale (n = 219), which measures resilience on a scale from 0 (no resilience) to 40 (greatest resilience), the mean score was 29.6 (SD, 6.9), with a median of 30 (IQR = 10). Intrinsic factors most strongly associated with greater resilience were increasing age (P = .01) and later career stage (P = .002). Job satisfaction, autonomy, good work-life balance, and approach-focused coping strategies were positively associated with resilience. Overwhelmingly, the primary reported barrier to performing healthy coping behaviors was limited time to devote to self-care (177/266 [67%]). CLINICAL RELEVANCE: A combination of individual approach-focused coping strategies and organizational interventions are crucial to support a resilient veterinary workforce.

Optimizing Pooled Testing for Estimating the Prevalence of Multiple Diseases.

Pooled testing can enhance the efficiency of diagnosing individuals with diseases of low prevalence. Often, pooling is implemented using standard groupings (2, 5, 10, etc.). On the other hand, optimization theory can provide specific guidelines in finding the ideal pool size and pooling strategy. This article focuses on optimizing the precision of disease prevalence estimators calculated from multiplex pooled testing data. In the context of a surveillance application of animal diseases, we study the estimation efficiency (i.e., precision) and cost efficiency of the estimators with adjustments for the number of expended tests. This enables us to determine the pooling strategies that offer the highest benefits when jointly estimating the prevalence of multiple diseases, such as theileriosis and anaplasmosis. The outcomes of our work can be used in designing pooled testing protocols, not only in simple pooling scenarios but also in more complex scenarios where individual retesting is performed in order to identify positive cases. A software application using the shiny package in R is provided with this article to facilitate implementation of our methods. Supplementary materials accompanying this paper appear online. Supplementary materials for this article are available at 10.1007/s13253-022-00511-4.

Distribution of Theileria orientalis in Virginia Market Cattle, 2018-2020.

Theileria orientalis, genotype Ikeda, was recently detected in North America. Determining the emerging distribution of this pathogen is critical for understanding spread and developing management strategies. Whole blood samples were collected from cattle at Virginia livestock markets from September 2018 through December 2020. Animals were tested for T. orientalis using a universal and then genotype specific real-time PCR based on the MPSP gene. Prevalence for each genotype was analyzed for temporal trends and mapped by county. Spatial patterns were compared between genotypes and assessed for associations with habitat features, cattle movements through cattle markets and county proximity. Overall, 212 of 1980 samples tested positive for T. orientalis with an overall prevalence of 8.7% (172/1980) for genotype Ikeda, 1.8% (36/1980) for genotype Chitose, 0.2% (3/1980) for genotype Buffeli. The Ikeda genotype increased over time in northern and southwestern Virginia markets. The Ikeda and Chitose genotypes occurred in different regions, with little overlap, but for each genotype, spatial distribution was associated with a combination of cattle movements and environmental factors. Genotype specific qPCR testing and surveillance of cattle from across a wide area of Virginia are providing information on temporal, spatial, and other patterns for this emerging disease.

Subsewershed SARS-CoV-2 Wastewater Surveillance and COVID-19 Epidemiology Using Building-Specific Occupancy and Case Data.

To evaluate the use of wastewater-based surveillance and epidemiology to monitor and predict SARS-CoV-2 virus trends, over the 2020-2021 academic year we collected wastewater samples twice weekly from 17 manholes across Virginia Tech's main campus. We used data from external door swipe card readers and student isolation/quarantine status to estimate building-specific occupancy and COVID-19 case counts at a daily resolution. After analyzing 673 wastewater samples using reverse transcription quantitative polymerase chain reaction (RT-qPCR), we reanalyzed 329 samples from isolation and nonisolation dormitories and the campus sewage outflow using reverse transcription digital droplet polymerase chain reaction (RT-ddPCR). Population-adjusted viral copy means from isolation dormitory wastewater were 48% and 66% higher than unadjusted viral copy means for N and E genes (1846/100 mL to 2733/100 mL/100 people and 2312/100 mL to 3828/100 mL/100 people, respectively; n = 46). Prespecified analyses with random-effects Poisson regression and dormitory/cluster-robust standard errors showed that the detection of N and E genes were associated with increases of 85% and 99% in the likelihood of COVID-19 cases 8 days later (incident-rate ratio (IRR) = 1.845, p = 0.013 and IRR = 1.994, p = 0.007, respectively; n = 215), and one-log increases in swipe card normalized viral copies (copies/100 mL/100 people) for N and E were associated with increases of 21% and 27% in the likelihood of observing COVID-19 cases 8 days following sample collection (IRR = 1.206, p < 0.001, n = 211 for N; IRR = 1.265, p < 0.001, n = 211 for E). One-log increases in swipe normalized copies were also associated with 40% and 43% increases in the likelihood of observing COVID-19 cases 5 days after sample collection (IRR = 1.403, p = 0.002, n = 212 for N; IRR = 1.426, p < 0.001, n = 212 for E). Our findings highlight the use of building-specific occupancy data and add to the evidence for the potential of wastewater-based epidemiology to predict COVID-19 trends at subsewershed scales.

Longitudinal social network analysis of avian mycobacteriosis incidence in a large population of zoo birds.

The goal of this study was to evaluate longitudinal patterns of avian mycobacteriosis spread through a social network. Specifically, we wanted to determine whether the patterns of connectivity over time can predict future infections, and whether this pattern can distinguish between different sources of infection. The study population included 13,409 individuals nested in a larger population of birds that were closely monitored in zoological facilities for over 22 years (1992-2014). A retrospective cohort study design and social network connectivity were used to estimate the association between exposure to an infected bird, and development of mycobacteriosis. Avian mycobacteriosis was diagnosed from histopathology and network connectivity was defined by enclosure histories over discrete time periods. Single-variable and multivariable longitudinal, mixed effects logistic regression models examined whether exposure to infected birds, both directly- and indirectly-connected, was associated with development of mycobacteriosis at the next time step. Our adjusted model showed an increased odds of developing mycobacteriosis (odds ratio = 2.15; 95 % CI: 1.48-3.12; p < 0.001) for birds that were directly exposed (i.e., housed in the same aviary) to another infected bird, compared to those with no exposure. Exposure to a positive, indirectly-connected bird at a previous time step was independently associated with an increased risk of mycobacteriosis (odds ratio = 1.56; 95 % CI: 1.11-2.19). This association persisted in adjusted models even when the indirect contacts were housed in distinctly different aviaries and never had contact with the subject of interest or its environment. Adjusted, risk-stratified models further characterized the type of exposure that increased the risk of avian mycobacteriosis. Birds that were exposed in small aviaries were more likely to develop mycobacteriosis than those exposed in larger aviaries and those with no exposure. The lesion distribution and species of the contact (same species versus different species) were also significant predictors of disease risk. Some findings were sensitive to model variation of time divisions and initiation time. Our study shows avian mycobacteriosis spread through the social network in quantifiable and discernable patterns. We provide empirical evidence that a contagious process drives some of the observed infection, but we also show low transmissibility based on sustained patterns of low incidence over time even when large groups of birds are exposed. Targeted risk mitigation efforts based on the characteristics of the exposure may be effective at reducing risk of avian mycobacteriosis while enhancing population sustainability.

Social network analysis and whole-genome sequencing to evaluate disease transmission in a large, dynamic population: A study of avian mycobacteriosis in zoo birds.

This study combined a social network analysis and whole-genome sequencing (WGS) to test for general patterns of contagious spread of a mycobacterial infection for which pathways of disease acquisition are not well understood. Our population included 275 cases diagnosed with avian mycobacteriosis that were nested in a source population of 16,430 birds at San Diego Zoo Wildlife Alliance facilities from 1992 through mid-2014. Mycobacteria species were determined using conventional methods and whole genome sequencing (WGS). Mycobacterium avium avium (MAA) and Mycobacterium genavense were the most common species of mycobacteria identified and were present in different proportions across bird taxa. A social network for the birds was constructed from the source population to identify directly and indirectly connected cases during time periods relevant to disease transmission. Associations between network connectivity and genetic similarity of mycobacteria (as determined by clusters of genotypes separated by few single nucleotide polymorphisms, or SNPs) were then evaluated in observed and randomly generated network permutations. Findings showed that some genotypes clustered along pathways of bird connectivity, while others were dispersed throughout the network. The proportion of directly connected birds having a similar mycobacterial genotype was 0.36 and significant (p<0.05). This proportion was higher (0.58) and significant for MAA but not for M. genavense. Evaluations of SNP distributions also showed genotypes of MAA were more related in connected birds than expected by chance; however, no significant patterns of genetic relatedness were identified for M. genavense, although data were sparse. Integrating the WGS analysis of mycobacteria with a social network analysis of their host birds revealed significant genetic clustering along pathways of connectivity, namely for MAA. These findings are consistent with a contagious process occurring in some, but not all, case clusters.

A retrospective analysis of bull:cow ratio effects on pregnancy rates of beef cows previously enrolled in fixed-time artificial insemination protocols.

This retrospective study aimed to determine if the number of cows exposed per bull affects pregnancy rates of cows returning to estrus after fixed-time artificial insemination (FTAI). Data were compiled over the course of 13 breeding seasons (six fall and seven spring seasons) between 2010 and 2017 from the Virginia Department of Corrections herd. Available records contained data from 17 farms and 324 groups of cows (average 47 cows/group). Multiparous cows and heifers (average age per group: 5.11 ± 0.14 yr; n = 14,868) were exposed to FTAI. After FTAI, animals were placed on pasture with bulls diagnosed as fertile by a breeding soundness exam for natural service of cows who did not become pregnant to FTAI (n = 7,248; average 22 cows/group). Animals were classified as pregnant to FTAI, to natural service on first return to estrus, or to natural service on second or subsequent estrus determined by fetal aging at pregnancy diagnosis. The bull:cow ratio for the total number of cows exposed ranged from 1:9 to 1:73 with an average of 1:31. The bull:cow ratio considering only open cows exposed after FTAI ranged from 1:2 to 1:44 with an average of 1:14. There was significant negative, small correlation between the bull:cow ratio for total number of cows exposed and return to estrus pregnancy rate in fall breeding seasons (P = 0.01, r 2 = 0.04) but not in spring (P = 0.90). There was a significant negative, small correlation between bull:cow ratio of open cows exposed and pregnancy rates to first return to estrus in fall herds with a single sire (P < 0.001, r 2 = 0.11). There was no correlation in fall herds using multiple sires or spring herds (P ≥ 0.12). Bull:cow ratio accounted for only 1-11% of variation in the pregnancy rates, thus we conclude that a decreased bull:cow ratio (up to 1:73) did not affect natural service return to estrus pregnancy rate. Cattlemen may consider a reduced number of bulls needed for natural service breeding after FTAI, which can decrease bull related costs and increase the economic feasibility of adopting FTAI protocols.

The impact of human reservoir of malaria at a community-level on individual malaria occurrence in a low malaria transmission setting along the Thai-Myanmar border.

BACKGROUND: The probability of contracting malaria in a given individual is determined not only by the individual's characteristics, but also the ecological factors that characterize the level of human-vector contact in the population. Examination of the relationship between "individual" and "supra-individual" variables over time is important for understanding the local malaria epidemiology. This is essential for planning effective intervention strategies specifically for each location. METHODS: A retrospective cohort study was conducted, which followed a community-cohort of about 3,500 residents in seven hamlets along the Thai-Myanmar border between 1999 and 2006. Potential malaria determinants measured at different levels (temporal variables, individual variables, and hamlet variables) were incorporated into multilevel models to estimate their effects on an individual's risk of malaria attack. RESULTS: The monthly minimum temperature was significantly associated with the seasonal variation of malaria risk. An individual risk of malaria attack decreased by about 50% during the period that active surveillance was conducted; an additional 15% and 25% reduction of Plasmodium falciparum and Plasmodium vivax incidence, respectively, was observed after the use of artesunate-mefloquine combination therapy (ACT) for treatment of P. falciparum. Male children (age < 16 years old) were at highest risk of both P. falciparum and P. vivax attack. An increase in the hamlet's incidence of P. falciparum and P. vivax by 1 per 100 persons in a previous month resulted in 1.14 and 1.34 times increase in the risk of P. falciparum and P. vivax, respectively, among individuals in a particular hamlet. CONCLUSION: In a small area with low malaria transmission intensity, the variation in mosquito abundance is relatively similar across the residential areas; incidence of malaria between hamlets, which reflects the community level of human infectious reservoirs, is an important predictor for the malaria risk among individuals within these hamlets. Therefore, local malaria control strategies should focus on interventions that aim to reduce the gametocyte carriage in the population, such as early detection and treatment programmes and the use of ACT for P. falciparum.

Investigation of factors predicting disease among zoo birds exposed to avian mycobacteriosis.

OBJECTIVE: To characterize infection patterns and identify factors associated with avian mycobacteriosis among zoo birds that were housed with infected enclosure mates. DESIGN: Matched case-control study. ANIMALS: 79 birds with avian mycobacteriosis (cases) and 316 nondiseased birds (controls) of similar age and taxonomic group that were present in the bird collection of the Zoological Society of San Diego from 1991 through 2005. PROCEDURES: Inventory and necropsy records from all eligible, exposed birds (n = 2,413) were examined to determine disease incidence and prevalence in the exposed cohort. Cases were matched in a 1:4 ratio to randomly selected controls of similar age and taxonomic grouping. Risk factors for mycobacteriosis (demographic, temporal, enclosure, and exposure characteristics as well as translocation history) were evaluated with univariate and multivariable conditional logistic regression analyses. RESULTS: Disease prevalence and incidence were estimated at 3.5% and 8 cases/1,000 bird-years at risk, respectively. In the multivariable model, cases were more likely to have been imported into the collection, exposed to mycobacteriosis at a young age, exposed to the same bird species, and exposed in small enclosures than were controls. Odds for disease increased with an increasing amount of time spent with other disease-positive birds. CONCLUSIONS AND CLINICAL RELEVANCE: The low incidence of mycobacteriosis and the risk factors identified suggested that mycobacteria may not be easily transmitted through direct contact with infected enclosure mates. Identification of risk factors for avian mycobacteriosis will help guide future management of this disease in zoo bird populations.