Bacterial burden and molecular characterization of Coxiella burnetii in shedding pregnant and postpartum ewes from Saint Kitts.

This study aimed to evaluate the bacterial burden and perform molecular characterization of Coxiella burnetii during shedding in pregnant (vaginal, mucus and feces) and postpartum (vaginal mucus, feces and milk) ewes from Saint Kitts. Positive IS1111 DNA (n=250) for C. burnetii samples from pregnant (n=87) and postpartum (n=74) Barbados Blackbelly ewes in a previous investigation were used for this study. Vaginal mucus (n=118), feces (n=100), and milk (n=32) positive IS1111 C. burnetii-DNA were analysed by real time qPCR (icd gene). For molecular characterization of C. burnetii, selected (n=10) IS1111 qPCR positive samples were sequenced for fragments of the IS1111 element and the 16 S rRNA gene. nBLAST, phylogenetic and haplotype analyses were performed. Vaginal mucus, feces and milk had estimated equal amounts of bacterial DNA (icd copies), and super spreaders were detected within the fecal samples. C. burnetii haplotypes had moderate to high diversity, were ubiquitous worldwide and similar to previously described in ruminants and ticks and humans.

In-nest mortality and pathology of hawksbill sea turtle (Eretmochelys imbricata) embryos and hatchlings in St. Kitts and Nevis.

Strategies to improve the hatch success and survival of critically endangered hawksbill turtles (Eretmochelys imbricata) require knowledge of threats to them including pathological conditions. The objective of this study was to describe the mortality and pathology of embryos and dead-in-nest hatchlings on St. Kitts and Nevis. Over the 2019/20 and 2020/21 nesting seasons, the combined mean (SD) hatch success for the two islands was 81.9% (13.2%) and deceased individuals within excavated nests were early-stage embryos (70.7%), late-stage embryos (17.7%), pipped-hatchlings (8.2%) or dead-in-nest hatchlings (3.4%). From 2017 to 2021, a post-mortem examination was performed on 183 turtles, including histology for 116. Anatomical malformations affected 77 (42.1%) examined turtles and included abnormal scute shape or number (22.4%), dysmelia (8.7%), schistosomus reflexus (7.7%) and compressed carapace (7.7%). Microscopic lesions were found in 49.1% of turtles and included tissue mineralization (26.7%, including renal, fetal membrane, liver, heart or muscle), chorioallantoitis (16.2%) and skeletal muscle degeneration and necrosis (10%). Inflammatory lesions associated with fungal or bacterial infections were in the skin (n = 3), chorioallantois (n = 4), lung (n = 3) or yolk sac (n = 1). These lesions may reflect non-specific terminal conditions but their presence in-nest helps explain some of the mortality and pathology documented in hatchlings that die during rehabilitative care. All of the gonads adequately represented for histological determination of sex were female (n = 62), supporting concern for feminization of Caribbean hawksbill turtle nests. The study identifies lesions that could affect hatch and emergence success. The high frequency of skeletal malformations indicates the need for investigations addressing regional impact and pathogenesis, especially genetic and environmental aetiologies including nest temperature. Immediate examination of live hatchlings on nest emergence is warranted to better determine the prevalence of non-fatal malformations that could impact fitness and population genetics.

Antemortem cytologic diagnosis of pulmonary Langerhans cell histiocytosis in a cat.

Feline pulmonary Langerhans cell histiocytosis (FPLCH) is a rare histiocytic proliferative disease of middle-aged to older domestic cats. Langerhans cells in the terminal airways proliferate and infiltrate the interstitium and the airways to a lesser degree, widely effacing normal parenchyma. Historically, definitive diagnosis has required postmortem evaluation where pulmonary lesions have a classic gross and histologic morphology. Here, we present the first documented antemortem diagnosis of FPLCH using bronchoalveolar lavage (BAL) cytology and immunocytochemistry (ICC) in a 9-year-old British shorthair mix. The cat had a 3-month history of respiratory difficulty that was refractory to steroids and antimicrobials. Pulmonary radiographs had marked diffuse changes with a complex bronchointerstitial and micronodular pattern. BAL cytology revealed neutrophilic inflammation and markedly increased histiocytes with morphology distinct from typical pulmonary macrophages. ICC characterized histiocytes as CD1a+ /E-cadherin+ /CD11b- /PanCK- , consistent with a Langerhans cell phenotype. The cat was humanely euthanized due to poor prognosis and presented for necropsy. Gross, histopathologic, immunophenotypic, and ultrastructural findings confirmed a diagnosis of FPLCH. Proliferative cells were E-cadherin+ /Iba-1+ /CD18+ /CD1a+ /CD5+ /MHCII+ /CD204- /CD4- ; transmission electron microscopy identified the presence of Birbeck granules in the proliferating histiocytes, consistent with previous reports of FPLCH.

Coxiella burnetii shedding and serological status in pregnant and postpartum ewes.

This study aimed to evaluate the occurence C. burnetii-DNA shedding by pregnant (vaginal mucus and feces) and postpartum (vaginal mucus, feces and milk) meat breed ewes from Saint Kitts. Additionally, antibodies anti-C. burnetii were detected in serum, and milk. Barbados Blackbelly ewes (n=187) were sampled using stratified convenience cross-sectional sampling. There were two animal groups: pregnant (n=96) and postpartum (n=91). Vaginal mucus (n=187), feces (n=177) and milk (n=83) samples were subjected to a TaqMan real time qPCR assay for C. burnetii based on the IS1111 multi copy element. IgG antibodies against C. burnetii were tested in blood serum (n=187) and milk (n=61) samples, via indirect ELISA. McNemar and Fischer exact tests were used to compare occurrence between routes and between groups, respectively. Overall, 86.6% of all the animals (162/187) were shedding C. burnetti DNA through at least one route (vaginal and/or fecal and/or milk). The DNA shedding occurrence via vaginal (73% vs 51%, p-value=0.003) and fecal routes (64% vs 47%, p-value=0.001) was higher in the pregnant compared to the postpartum animals. There was no prevalent shedding route among vaginal, fecal or milk in all ewes. Overall, 38% of the ewes were seropositive for C. burnetii IgG and a total of 19.7% of the tested postpartum ewes had IgG antibodies in milk. The vaginal and fecal DNA shedding were not associated with the blood serology, nor was milk DNA shedding related to the milk serology status, thus there was no association between C. burnetii seropositivity and bacterial DNA shedding. In short, high occurrence of C. burnetii DNA shedding was observed within ewes in St. Kitts, and represents the first detection of the Q fever agent within the Caribbean islands. Bacterial shedding was more prevalent in pregnant ewes, highlighting the importance of gestating animals as a source of C. burnetii.

Molecular Survey and Genetic Diversity of Bartonella spp. in Small Indian Mongooses (Urva auropunctata) and Their Fleas on Saint Kitts, West Indies.

This study aimed to molecularly survey and evaluate the genetic diversity of Bartonella spp. in mongooses and their fleas from St. Kitts. Spleen (n = 54), blood (n = 71), and pooled flea samples, all identified as Ctenocephalides felis (n = 53), were submitted to TaqMan real-time quantitative PCR (qPCR) targeting Bartonella-nuoG fragment (84 bp). Positive samples underwent further conventional PCR assays targeting five loci (gltA, rpoB, fstZ, nuoG, and ITS), subsequent sequencing, and phylogenetic and haplotype analyses. The overall occurrence of Bartonella spp. in mongooses and fleas was 51.2% (64/125 [95% CI (42.1-60.2%)]) and 62.3% (33/53) [95% CI (47.9-75.2%)]), respectively. From samples sequenced across the five loci, 50.8% (33/65) were identified as Bartonella henselae, 26.2% (17/65) were 96.74-99.01% similar by BLAST analysis to an unidentified Bartonella sp. previously reported in Japanese badgers (Meles anakuma), and 23.1% (15/65) were co-infected with both species. Nucleotide polymorphism analysis showed low diversity amongst haplotypes but did concur with phylogenetic analysis, placing the unidentified species in a separate clade from B. henselae by multiple mutational events. Our data confirms that mongooses and Ctenocephalides felis fleas collected from them are not only potential reservoirs for B. henselae but also a novel Bartonella sp. which we propose be called 'Candidatus Bartonella kittensis'.