Brucella melitensis: Divergence Among Indian Strains and Genetic Characterization of a Strain Isolated from Cattle.

Brucella melitensis primarily affects sheep, goats and is associated with brucellosis in humans, which is one of the world's most widespread neglected zoonotic disease. The current study attempted the determination of genetic diversity through comparative genome analysis of B. melitensis strains reported from India with other countries. The study also reports the isolation and identification of B. melitensis BMNDDB8664 from a cow with a history of abortion, whole-genome sequencing (WGS), determination of virulence factors, genotyping, and comparative genome analysis. Multilocus sequence typing, Multiple locus variable number of tandem repeats analysis (MLVA), and WGS based phylogeny revealed the predominance of ST-8 and genotypes (116 and II respectively) that clustered to the East Mediterranean lineage. Identification of hitherto unreported genotypes by MLVA also indicated the existence and circulation of West Mediterranean and American lineages in India. Though the AMOS-PCR results suggest the BMNDDB8664 isolate as Brucella abortus, the outcomes from multiplex PCR, ribosomal multilocus sequence typing, and WGS analysis confirmed it as B. melitensis. The analysis revealed the presence of adeF gene (aids conferring resistance to fluoro-quinolone and tetracyclines). The isolate lacked two important T4SS genes virB2 and virB7 genes (roles in infection and rifampicin resistance respectively) and also lacked the Brucella suis mprF gene that aids intracellular survival. Further, BMNDDB8664 lacked some of the genes associated with LPS synthesis (wbkB, wbkC) and transport (wzm, wzt) and hence, is most likely a rough strain. WGS-based phylogenetic analysis revealed close genetic relatedness of this BMNDDB8664 with a sheep isolate and two human isolates. The results prompt systematic, broad-based epidemiological studies on brucella infection at the species level. For effective control of human brucellosis, a concerted One Health approach with studies encircling the identification of aetiology at species, strain level to find their prevalence, spread, and inter-host transmission patterns need to be understood, for better design and implementation of effective control strategies in India and other endemic regions. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-023-01081-w.

Seroprevalence of Crimean-Congo haemorrhagic fever in Indian cattle and buffaloes.

BACKGROUND & OBJECTIVES: Crimean-Congo hemorrhagic fever (CCHF) is an emerging tick-borne viral zoonotic disease of public health importance. Cattle and buffaloes although not showing any clinical symptoms, can be infected by the CCHF virus and act as sources of infection to human beings. The prevalence of CCHF in cattle and buffaloes is important from One health perspective for control of CCHF in humans. METHODS: A cross-sectional study was undertaken to ascertain the prevalence of CCHFV in cattle and buffaloes of India. MATERIALS AND METHODS: A total of 804 serum samples from four states of India (Gujarat and Rajasthan: human outbreaks reported; Punjab and Haryana: no outbreak reported) were screened by ELISA test detecting nucleoprotein antibodies of CCHFV. RESULTS: The overall true prevalence was 8.63% (95% CI: 6.76% - 10.9%). The highest prevalence was recorded in Rajasthan (13.24%) followed by Gujarat (8.68%), Haryana (6.84%), and Punjab (6.51%). Prevalence of CCHF was higher in cattle (9.92%) than buffaloes (5.84%); in females (10.87%) than males (4.99%); in adults (10.18%) than young ones (5.66%). Interestingly, higher seropositivity was recorded in indigenous cattle (12.04%) than in exotic and cross-breed cattle (1.69%) which was statistically significant (p=0.001). INTERPRETATION & CONCLUSION: These findings revealed CCHF virus is circulating unnoticed and the prevalence has increased over time which is of public health concern.

Detection and genetic characterization indicates circulation of a possible new Theileria species (Theileria sp. Yokoyama) in India.

Bovine tropical theileriosis, a tick-borne disease, causes huge economic loss to the Indian dairy industry. Theileriosis in India is mainly caused by Theileria annulata, although the presence of T. orientalis has also been reported. The present study was undertaken to investigate the deaths of cross-bred Holstein Friesen (CBHF) cows on a farm in the state of Telangana, India. Deceased animals had recently calved and prior to death had developed high fever (107 °F) and anaemia. Infected cows were infested with ticks (Hyalomma species). Theileria piroplasms were noticed in the Giemsa stained blood smears. PCR assays further confirmed the presence of Theileria in the blood samples of the infected cows. Partial Tams1 gene sequences from the infected animals shared 99.87% to 100% identity scores with the sequences of Sri Lankan isolates recently proposed as a novel Theileria species (provisionally designated as Theileria sp. Yokoyama). To the best of our knowledge, this is the first report of the novel species of Theileria from India. Infected animals were effectively treated with buparvaquone and oxytetracycline. The introduction of new animals into the farm without risk assessment was found to be a major cause of the outbreak.

Attributable mortality and excess length of stay associated with third-generation cephalosporin-resistant Enterobacterales bloodstream infections: a prospective cohort study in Suva, Fiji.

OBJECTIVES: There are scant primary clinical data on antimicrobial resistance (AMR) burden from low- and middle-income countries (LMICs). We adapted recent World Health Organization methodology to measure the effect of third-generation cephalosporin resistance (3GC-R) on mortality and excess length of hospital stay in Fiji. METHODS: We conducted a prospective cohort study of inpatients with Enterobacterales bloodstream infections (BSIs) at Colonial War Memorial Hospital, Suva. We used cause-specific Cox proportional hazards models to estimate the effect of 3GC-R on the daily risk (hazard) of in-hospital mortality and being discharged alive (competing risks), and we used multistate modelling to estimate the excess length of hospital stay. RESULTS: From July 2020 to February 2021 we identified 162 consecutive Enterobacterales BSIs; 3GC-R was present in 66 (40.7%). Crude mortality for patients with 3GC-susceptible and 3GC-R BSIs was 16.7% (16/96) and 30.3% (20/66), respectively. 3GC-R was not associated with the in-hospital mortality hazard rate (adjusted hazard ratio [aHR] 1.13, 95% confidence interval [CI] 0.51-2.53) or being discharged alive (aHR 0.99, 95% CI 0.65-1.50), whereas Charlson comorbidity index score (aHR 1.62, 95% CI 1.36-1.93) and Pitt bacteraemia score (aHR 3.57, 95% CI 1.31-9.71) were both associated with an increased hazard rate of in-hospital mortality. 3GC-R was associated with an increased length of stay of 2.6 days (95% CI 2.5-2.8). 3GC-R was more common among hospital-associated infections, but genomics did not identify clonal transmission. CONCLUSION: Patients with Enterobacterales BSIs in Fiji had high mortality. There were high rates of 3GC-R, which was associated with increased hospital length of stay but not with in-hospital mortality.

Epidemiology, antimicrobial resistance and outcomes of Staphylococcus aureus bacteraemia in a tertiary hospital in Fiji: A prospective cohort study.

Background: Staphylococcus aureus bacteraemia (SAB) is one of the commonest bloodstream infections globally and is associated with a high mortality rate. Most published data comes from temperate, high-income countries. We describe the clinical epidemiology, microbiology, management and outcomes of patients with SAB treated in a tropical, middle-income setting at Fiji's largest hospital. Methods: A prospective, observational study was performed of consecutive SAB cases admitted to Colonial War Memorial Hospital (CWMH) in Suva, between July 2020 and February 2021. Detailed demographic, clinical and microbiological data were collected, including the key outcome of in-patient mortality. To estimate the population incidence, all SAB cases diagnosed at the CWMH laboratory were included - even if not admitted to CWMH - with the population of Fiji's Central Division used as the denominator. Findings: A total of 176 cases of SAB were detected over eight-months, which equated to an incidence of 68.8 cases per 100,000 population per year. Of these, 95 cases were admitted to CWMH within 48 h of index culture. Approximately 8.4% (8/95) of admitted cases were caused by methicillin-resistant Staphylococcus aureus (MRSA). All cause in-patient mortality was 25.3%, increasing to 55% among patients aged 60 or older. Interpretation: This reported incidence of SAB in central Fiji is one of the highest in the world. SAB was associated with significant mortality, especially in those over 60 years of age, despite a relatively low frequency of methicillin resistance. Funding: Supported by the National Health and Medical Research Council (Australia) and the GRAM (Global Research on Antimicrobial Resistance) Project, Oxford University (United Kingdom).

Evaluation of commercial ELISA kits for diagnosis of brucellosis in cattle and buffaloes in different epidemiological scenarios.

Seven ELISA kits were evaluated for the fitness of purpose in diagnosing brucellosis among cattle and buffaloes in the endemic scenarios of India. The sera (675 numbers) for the study were sourced from brucellosis-free as well as infected herds. The diagnostic sensitivity (dsn) and specificity (dsp) of the kits were determined by three approaches: based on the results of the Rose Bengal test, history of the animals (sera from infected or naïve animals), and based on the results obtained from the 'majority of the tests'. The dsn and dsp ranged from 65.10% to 98.66%, and 98.04% to 100% respectively. The results and suitability of the kits for diagnostic application in various epidemiological situations were discussed.

Prevalence of antibodies to Anaplasma in cattle and buffaloes of different organized herds in India.

Bovine anaplasmosis is one of the most important tick borne disease in ruminants causing huge economic loss to the dairy industry. A cross-sectional study was carried out to detect serum antibodies to Anaplasma infection in cattle and buffaloes housed in 14 organized herds located at various climatic zones spreading over 9 different states in India. A total of 911 serum samples, collected from 667 cattle and 244 buffaloes, were subjected to a competitive enzyme linked immune-sorbent assay detecting an epitope of major surface protein 5 (MSP5) of Anaplasma. The overall true prevalence was 48.72% (95% CI 45.13-52.32%). The prevalence rate was higher in cattle (51.58%) than buffaloes (40.89%) and the difference was statistically significant (p < 0.05). Indigenous cattle (59.30%) showed higher seropositivity than crossbreed (57.16%) and exotic cattle breeds (42.28%). Although statistically not significant, female (52.37%) showed higher seropositivity than male (46.43%). Similarly, significant difference in prevalence (p < 0.05) was observed for animals reared in different climatic zones with highest prevalence recorded in arid zone (90.49%) and lowest in semi-arid zone (29.83%). Very wide variation in prevalence (9.95-100%) was recorded between farms. The present study indicates endemicity of Anaplasma in India, similar to other tropical and sub-tropical countries of the world. Endemic instability was recorded in some of the studied farms suggesting possibility of outbreak of new clinical cases resulting in economic loss. Therefore, suitable policies and procedures for prevention and control of Anaplasma infection should be adopted in these farms.

Infectious bovine abortions: observations from an organized dairy herd.

Abortions in dairy animals can be caused by several infectious agents. Identification of the actual causal agent(s) is important for formulating suitable control strategies. A 3-year (2016-2018) longitudinal study was conducted in a dairy farm following an abortion storm in the mid- to late gestations. The investigation focused on the seven major infectious abortifacient in cattle, viz. bovine alphaherpesvirus-1 (BoHV-1), bovine viral diarrhoea virus (BVDV), Neospora caninum, Brucella abortus, Coxiella burnetii, Leptospira Hardjo, and Listeria monocytogenes. High seroprevalence was observed for BVDV (79.4%), Leptospira (70.5%), BoHV-1 (53.5%), and Brucella (45.0%) at the beginning of the investigation (August 2016). The incidence proportion increased for BVDV, Leptospira, and Brucella in the following years of the investigation. A strong association of Brucella seropositivity with history of abortion (OR = 3.27) was recorded. Incidence of BoHV-1 reduced during the period of study coincident with systematic IBR inactivated marker vaccination of the herd. Sixty-four abortion cases were investigated for the identification of causative agent(s) by microbial culture, serological (ELISA), and molecular detection (PCR/ real-time PCR). Antibodies to BVDV, Brucella, BoHV-1, Leptospira, Neospora, and Coxiella were detected in 63, 61, 56, 35, 5, and 6 aborting cattle, respectively. Real-time PCR/PCR of clinical specimens detected DNA of Brucella, BoHV-1, Coxiella, Leptospira, and Listeria in 34, 13, 12, 9, and 4 abortion cases, respectively. BVDV and Neospora were not detected in any specimen samples. Brucella abortus isolated from the farm was determined as ST1 by multi-locus sequence typing (MLST). DNA of multiple agents were detected in 21 of the 64 cases (43.75%). Overall, the data suggests, Brucella was the major causative agent, although multiple causative agents circulated in the farm.

What is the prevalence of senior-athlete rotator cuff injuries and are they associated with pain and dysfunction?

BACKGROUND: Older individuals with rotator cuff injuries may have difficulties not only with activities of daily living, but also with sports activities. QUESTIONS/PURPOSES: (1) How frequent and severe are rotator cuff abnormalities, as identified by ultrasound, in senior athletes? (2) To what degree does the severity of ultrasound-identified rotator cuff pathology correlate with pain and shoulder dysfunction? METHODS: We assessed pain and shoulder function in 141 elite athletes older than 60 years of age (median age, 70 years; range 60-84) at the Senior Olympics who volunteered to participate. An ultrasound evaluation of the rotator cuff of the dominant shoulder was performed by an experienced musculoskeletal radiologist in all of these elite athletes. We then determined the relationship between ultrasound findings and shoulder pain and shoulder function as assessed with the Disabilities of the Arm, Shoulder and Hand (DASH) and American Shoulder and Elbow Surgeons (ASES) scores. RESULTS: There were 20 shoulders with a normal cuff (14.2% [20 of 141], of which 5% [one of 20] were painful), 23 with tendinosis (16.3% [23 of 141], of which 30% [six of 20] were painful), 68 with a partial-thickness rotator cuff tear (48.2% [68 of 141], of which 32% [20 of 63] were painful), and 30 with a full-thickness rotator cuff tear (21.3% [30 of 141], of which 25% [seven of 28] were painful). Only 5% of athletes (one of 20) with a normal cuff on ultrasound evaluation reported shoulder pain, whereas 30% of athletes (33 of 111) with any degree of rotator cuff damage on ultrasound evaluation reported shoulder pain, This resulted in an odds ratio of 8.0 (95% confidence interval, 1.0-62.5). The proportion of patients who had pain was not different in those with different severities of rotator cuff pathology. Neither the ASES nor the DASH was different in those with different severities. CONCLUSIONS: The frequency of full-thickness rotator cuff tears in senior athletes was 21.3% (30 of 141). Pain was a predictor of rotator cuff injury but not of its severity. The odds of having shoulder pain was eight times greater in those athletes with any rotator cuff damage as compared with those without any rotator cuff damage. Those with pain had poorer shoulder function but the ASES and DASH were poor predictors of the severity of rotator cuff pathology. Rotator cuff tears in older individuals are often not painful and may not need to be repaired for successful participation in athletics. LEVEL OF EVIDENCE: Level II, prognostic study. See Guidelines for Authors for a complete description of levels of evidence.

Full-thickness and partial-thickness supraspinatus tendon tears: value of US signs in diagnosis.

PURPOSE: To determine which US signs are important in the diagnosis of a surgically identifiable supraspinatus tendon tear. MATERIALS AND METHODS: Fifty consecutive ultrasonographic (US) studies of the shoulder in patients who underwent arthroscopic follow-up were retrospectively reviewed by a musculoskeletal radiologist. US images of the supraspinatus tendon were evaluated for tendon nonvisualization, abnormal tendon echogenicity, tendon thinning, greater tuberosity cortical irregularity, cartilage interface sign, joint fluid, and subacromial-subdeltoid bursal fluid. US findings were compared with arthroscopic results. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were calculated for each US sign in the diagnosis of full-thickness tendon tear and again for any type of supraspinatus tendon tear. RESULTS: Arthroscopy revealed 21 full-thickness tears, five bursal surface partial-thickness tears, 10 articular surface partial-thickness tears, and 14 patients without tear of the supraspinatus tendon. The presence of greater tuberosity cortical irregularity and joint fluid was most important in the diagnosis of full-thickness supraspinatus tendon tear (sensitivity, 60%; specificity, 100%; positive predictive value, 100%; negative predictive value, 78%; accuracy, 84%). For diagnosis of any type of supraspinatus tendon tear (partial or full thickness), tendon nonvisualization, greater tuberosity cortical irregularity, and cartilage interface sign are most important, although a combination of signs did not improve accuracy. CONCLUSION: Secondary US signs, such as greater tuberosity cortical irregularity and joint fluid, are most valuable in the diagnosis of supraspinatus tendon tear.

Elbow: the trochleogingylomoid joint.

More complex than a hinged joint, the elbow provides flexion-extension as well as pronation-supination. A unique arrangement of joint surfaces, soft tissue restraints, and muscle forces provides the mobility and stability of this joint. Understanding basic elbow biomechanics will assist the musculoskeletal radiologist in evaluations of this joint.