A model for chronic bovine besnoitiosis: Parasite stage and inoculation route are key factors

In this work, an experimental model for chronic besnoitiosis in bovine was developed and characterized. Using a previously established calf model, two new variables (parasite stage and inoculation route) were combined and used. Twelve Holstein Friesian 3‐month‐old male calves were randomly divided into four groups of three animals each. Bradyzoites were obtained from a chronically infected bull and used for inoculation via three different inoculation routes. Three groups were inoculated with 10⁶ bradyzoites by intravenous (G1), subcutaneous (G2) and intradermal (G3) routes, and a non‐infected control group (G4) was inoculated with PBS. The trial lasted for 90 days and included daily clinical monitoring as well as weekly skin biopsies and blood sampling. Sera were obtained to analyse both cellular and humoral responses. Once the calves were euthanized, tissues from the skin, eyes, respiratory and reproductive tracts, among others, were collected to study presence of the parasite. Clinically, the infection was classified as mild to moderate for the acute stage since all infected calves showed lymphadenopathy from four days post‐infection (pi) and fever from one week pi until 24 days pi. However, the most relevant results were achieved during the chronic stage that was classified as moderate to severe. In fact, pathognomonic conjunctival cysts were observed in all infected calves from 40 days pi onwards and were more abundant in G3. Moreover, one calf from this group developed skin lesions (49 days pi). The microscopic tissue cysts and Besnoitia DNA were detected primarily in skin, reproductive tract and respiratory tissue samples, and parasite load was higher in G3. In conclusion, the parasite stage (bradyzoite) and the inoculation route are key factors that influence the outcome of an infection. In particular, the intradermal route led to more severe clinical signs of the chronic phase in the inoculated calves.     

Clinical and Serological Dynamics of Besnoitia besnoiti Infection in Three Endemically Infected Beef Cattle Herds

The dynamics of bovine besnoitiosis were studied in an area where the disease is endemic. A four‐year longitudinal study was conducted for the first time in three infected beef cattle herds located in the Urbasa‐Andía Mountains (Navarra, Spain). Each herd was visited four to seven times, and clinical and serological prevalence rates and incidence rates were estimated. Clinical inspections to identify compatible clinical signs with the disease stages were conducted at the beginning and end of the study. Serological assessment was initially performed by ELISA. Seronegative animals with clinical signs and seropositive animals with relative index per cent (RIPC) values lower than 30 that did not increase during the study period were analysed by Western blot to optimize the sensitivity and specificity of the ELISA test. Clinical prevalence rates were slightly higher (62% on average) than the seroprevalence rates (50% on average), and tissue cysts located in the vestibulum vaginae and sclera were the most frequently detected clinical signs. The proportion of seropositive animals with clinical signs varied from 16.7% to 73.6% among the herds, and 17% of cattle with clinical signs proved to be seronegative by both serological tests. An average 22% serological incidence rate was also reported in addition to clinical incidence rates that varied from 12.5% to 16.7%. Additionally, parasitemia was investigated in the herd that showed the highest clinical and seroprevalence rates. Only one PCR positive blood sample was detected. Thus, the role that blood may play in parasite transmission needs to be further investigated. Infected herds maintained both high prevalence and incidence rates in the absence of control measures and a high number of parasite carriers. Finally, economic impact studies on reproductive and productive losses associated with besnoitiosis need to be performed to implement a cost–benefit control programme.     

Liver fluke (Fasciola hepatica) co‐infection with bovine tuberculosis (bTB) in cattle: A retrospective animal‐level assessment of bTB risk in dairy and beef cattle

Bovine tuberculosis (bTB), caused by Mycobacterium bovis, remains a persistent problem for cattle industries in endemic countries. The frequency, quality, and performance of tests, and the presence of wildlife reservoirs, have been identified as impediments to eradication. Recently, exposure to helminth infection (Fasciola hepatica) has been associated negatively with the disclosure of bTB. Here, for the first time, we assess impact of concurrent infections of Fasciola hepatica and the disclosure of bTB at the animal‐level using large surveillance datasets. We utilized a dataset of 138,566 animal records from an abattoir from Northern Ireland (2011–2013). The presence of F. hepatica infection was assessed from macroscopic tissue inspection at abattoir. Multivariable models were developed to assess co‐infection associations with bTB status based on: Single Intradermal Comparative Tuberculin Test (SICTT), lesion, bacteriological confirmation, including either all animals, or only skin‐test negative animals (lesions at routine slaughter; LRS; confirmed nonreactors at routine slaughter; cNRs) or positive (reactors) animals alone, respectively. The relationship between skin tuberculin reaction sizes and fluke status was also explored for a subset of animals with field recordings (n = 24,680). Controlling for known risk factors (e.g., climatic, herd, and individual level characteristics), we did not find significant associations between the SICTT (standard or severe interpretation), lesion, nor confirmation status of animals and their liver fluke status. The only exception was a negative association between liver fluke positivity, and LRS or cNRs, respectively; though effect‐sizes were small (e.g., LRS Odds‐Ratio: 0.87; 95% CI: 0.76–1.00). There was limited evidence of a relationship between tuberculin reaction sizes during SICTT testing and liver fluke infection status. These data do not support the contention that the detection of bTB using skin‐tests or reactor postmortem follow‐up may be compromised by co‐infection at a population level, but the relationship with lesion formation (pathogenesis) may indicate an impact for postmortem surveillance.     

Treatment preferences and outcome in male hypogonadotropic hypogonadism: an Indian perspective

This retrospective study assessed treatment preferences and outcome with testosterone or HCG / HCG–FSH combination in Indian male idiopathic hypogonadotropic hypogonadism (IHH) subjects (n = 31) above 18 years of age. 38.7% of IHH study subjects had no fertility plans and chose 3 monthly intramuscular testosterone undecanoate. 73.7% of subjects with fertility plans chose human chorionic gonadotropin (HCG) alone due to cost considerations. Spermatogenesis occurred in 21.4% on HCG alone and 60% of subjects on HCG with follicle‐stimulating hormone (FSH) combination. Treatment failure is higher than published Western rates. FSH and HCG combination regimen is costly but superior to HCG alone. However, treatment failure still persists, suggesting unknown testicular defect in IHH.     

Liver fluke (Fasciola hepatica) co‐infection with bovine tuberculosis in cattle: A prospective herd‐level assessment of herd bTB risk in dairy enterprises

Co‐infection of tuberculosis (TB) and helminths is recognized as a significant problem in regions where such pathogens are endemic and chronic cases exist. Co‐infection can modulate the immune system leading to interference with diagnostic tests, increased pathological impacts and pathogen persistence. However, research has found that such interactions between pathogens can be context and species specific. Recent studies have suggested that liver fluke, Fasciola hepatica, infection may impact on immunological responses and diagnostics for bovine tuberculosis (bTB; caused by Mycobacterium bovis) in cattle. Where evidence of such interaction exists, there would be an onus on policy makers to adjust eradication programs to minimize impacts. We assessed the association between herd‐level bTB breakdown risk and seasonal variation in liver fluke exposure based on 5,753 bulk tank milk (BTM) samples from 1,494 dairy herds across Northern Ireland. BTM was tested by an IDEXX antibody specific enzyme‐linked immunosorbent assay (ELISA) using the ‘f2’ antigen as a detection agent. The ELISA determined the result based on a sample to (known) positive ratio (S/P%) from which binary status and categories of exposure were derived. Associations were tested using multivariable random effects models. Models predicting bTB risk were not improved with the inclusion of liver fluke exposure levels. Variations in modelling liver fluke exposure (S/P%, binary, categories of exposure) and bTB risk (skin test breakdowns, post‐mortem confirmed breakdowns, breakdown size and lag effects) also failed to support associations (neither positive nor negative) between the pathogens at herd‐level. These results, along with previously published animal‐level data from Northern Ireland, suggest that the nexus between bTB and F. hepatica may have small size effects at the population‐level. However, our results also highlight the high prevalence of F. hepatica in cattle in our study population, and therefore we cannot fully discount the potential hypothesis of population‐level depression of immune response to M. bovis due to co‐infection.     

Disease Control in Wildlife: Evaluating a Test and Cull Programme for Bovine Tuberculosis in African Buffalo

Providing an evidence base for wildlife population management is difficult, due to limited opportunities for experimentation and study replication at the population level. We utilized an opportunity to assess the outcome of a test and cull programme aimed at limiting the spread of Mycobacterium bovis in African buffalo. Buffalo act as reservoirs of M. bovis, the causative agent of bovine tuberculosis (BTB), which can have major economic, ecological and public health impacts through the risk of infection to other wildlife species, livestock and surrounding communities. BTB prevalence data were collected in conjunction with disease control operations in Hluhluwe–iMfolozi Park, South Africa, from 1999 to 2006. A total of 4733 buffalo (250–950 per year) were tested for BTB using the single comparative intradermal tuberculin (SCIT) test, with BTB‐positive animals culled, and negative animals released. BTB prevalence was spatially and temporally variable, ranging from 2.3% to 54.7%. Geographic area was a strong predictor of BTB transmission in HiP, owing to relatively stable herds and home ranges. Herds experiencing more intensive and frequent captures showed reduced per capita disease transmission risk and less increase in herd prevalence over time. Disease hot spots did not expand spatially over time, and BTB prevalence in all but the hot spot areas was maintained between 10% and 15% throughout the study period. Our data suggest that HiP's test and cull programme was effective at reducing BTB transmission in buffalo, with capture effort and interval found to be the crucial components of the programme. The programme was thus successful with respect to the original goals; however, there are additional factors that should be considered in future cost/benefit analyses and decision‐making. These findings may be utilized and expanded in future collaborative work between wildlife managers, veterinarians and scientists, to optimize wildlife disease control programmes and mitigate conflict at the interface of conservation, agricultural and urban areas.     

Evidence of synergistic relationships between HIV and Human Papillomavirus (HPV): systematic reviews and meta‐analyses of longitudinal studies of HPV acquisition and clearance by HIV status,and of HIV acquisition by HPV status

Introduction

Observational studies suggest HIV and human papillomavirus (HPV) infections may have multiple interactions. We reviewed the strength of the evidence for the influence of HIV on HPV acquisition and clearance, and the influence of HPV on HIV acquisition.

Methods

We performed meta‐analytic systematic reviews of longitudinal studies of HPV incidence and clearance rate by HIV status (review 1) and of HIV incidence by HPV status (review 2). We pooled relative risk (RR) estimates across studies using random‐effect models. I² statistics and subgroup analyses were used to quantify heterogeneity across estimates and explore the influence of participant and study characteristics including study quality. Publication bias was examined quantitatively with funnel plots and subgroup analysis, as well as qualitatively.

Results and Discussion

In review 1, 37 publications (25 independent studies) were included in the meta‐analysis. HPV incidence (pooled RR = 1.55, 95% CI: 1.29 to 1.88; heterosexual males: pooled RR = 1.95, 95% CI: 1.62, 2.34; females: pooled RR = 1.63, 95% CI: 1.26 to 2.11; men who have sex with men: pooled RR = 1.36, 95% CI: 1.01 to 1.82) and high‐risk HPV incidence (pooled RR = 2.20, 95% CI: 1.90 to 2.54) was approximately doubled among people living with HIV (PLHIV) whereas HPV clearance rate (pooled RR = 0.53, 95% CI: 0.42 to 0.67) was approximately halved. In review 2, 14 publications (11 independent studies) were included in the meta‐analysis. HIV incidence was almost doubled (pooled RR = 1.91, 95% CI 1.38 to 2.65) in the presence of prevalent HPV infection. There was more evidence of publication bias in review 2, and somewhat greater risk of confounding in studies included in review 1. There was some evidence that adjustment for key confounders strengthened the associations for review 2. Misclassification bias by HIV/HPV exposure status could also have biased estimates toward the null.

Conclusions

These results provide evidence for synergistic HIV and HPV interactions of clinical and public health relevance. HPV vaccination may directly benefit PLHIV, and help control both HPV and HIV at the population level in high prevalence settings. Our estimates of association are useful for mathematical modelling. Although observational studies can never perfectly control for residual confounding, the evidence presented here lends further support for the presence of biological interactions between HIV and HPV that have a strong plausibility.

     

HIV incidence among people who inject drugs in the Middle East and North Africa: mathematical modelling analysis

Introduction

Emerging HIV epidemics have been documented among people who inject drugs (PWID) in the Middle East and North Africa (MENA). This study estimates the HIV incidence among PWID due to sharing needles/syringes in MENA. It also delineates injecting drug use role as a driver of the epidemic in the population, and estimates impact of interventions.

Methods

A mathematical model of HIV transmission among PWID was applied in seven MENA countries with sufficient and recent epidemiological data and HIV prevalence ≥1% among PWID. Estimations of incident and/or prevalent infections among PWID, ex‐PWID and sexual partners of infected current and ex‐PWID were conducted.

Results

The estimated HIV incidence rate for 2017 among PWID ranged between 0.7% per person‐year (ppy) in Tunisia and 7.8% ppy in Pakistan, with Libya being an outlier (24.8% ppy). The estimated number of annual new infections was lowest in Tunisia (n = 79) and Morocco (n = 99), and highest in Iran and Pakistan (approximately n = 6700 each). In addition, 20 to 2208 and 5 to 837 new annual infections were estimated across the different countries among sexual partners of PWID and ex‐PWID respectively. Since epidemic emergence, the number of total ever acquired incident infections across countries was 706 to 90,015 among PWID, 99 to 18,244 among sexual partners of PWID, and 16 to 4360 among sexual partners of ex‐PWID. The estimated number of prevalent infections across countries was 341 to 23,279 among PWID, 119 to 16,540 among ex‐PWID, 67 to 10,752 among sexual partners of PWID, and 12 to 2863 among sexual partners of ex‐PWID. Increasing antiretroviral therapy (ART) coverage to the global target of 81% – factoring in ART adherence and current coverage – would avert about half of new infections among PWID and their sexual partners. Combining ART with harm reduction could avert over 90% and 70% of new infections among PWID and their sexual partners respectively.

Conclusions

There is considerable HIV incidence among PWID in MENA. Of all new infections ultimately due to injecting drug use, about 75% are among PWID and the rest among sexual partners. Of all prevalent infections ultimately attributed to injecting drug use as epidemic driver, about half are among PWID, 30% among ex‐PWID and 20% among sexual partners of PWID and ex‐PWID. These findings call for scale‐up of services for PWID, including harm reduction as well as testing and treatment services.

     

Genetically ‘pure’ Fasciola gigantica discovered in Algeria: DNA multimarker characterization,trans‐Saharan introduction from a Sahel origin and spreading risk into north‐western Maghreb countries

Fascioliasis is a freshwater snail‐borne zoonotic helminth disease caused by two species of trematodes: Fasciola hepatica of almost worldwide distribution and the more pathogenic F. gigantica restricted to parts of Asia and most of Africa. Of high pathological impact in ruminants, it underlies large livestock husbandry losses. Fascioliasis is moreover of high public health importance and accordingly included within the main neglected tropical diseases by WHO. Additionally, this is an emerging disease due to influences of climate and global changes. In Africa, F. gigantica is distributed throughout almost the whole continent except in the north‐western Maghreb countries of Morocco, Algeria and Tunisia where only F. hepatica is present. The present study concerns the DNA multimarker characterization of the first finding of F. gigantica in sheep in Algeria by the complete sequences of rDNA ITS‐1 and ITS‐2 and mtDNA cox1 and nad1 genes. Sequence comparisons and network analyses show sequence identities and similarities suggesting a south–north trans‐Saharan geographical origin, with introduction from Ghana, through the Sahel countries of Burkina Faso and Mali into Algeria. This way perfectly fits with nomadic pastoralism according to interconnecting intranational and transborder herd transhumance routes traditionally followed in this western part of Africa from very long ago. The risk for further spread throughout the three north‐western Maghreb countries is multidisciplinarily analysed, mainly considering the present extensive motorization of the intranational transhumance system in Algeria, the lymnaeid snail vector species present throughout the north‐western Maghreb, the increasing demand for animal products in the growing cities of northern Algeria and the continued human infection reports. Control measures should assure making anti‐fasciolid drugs available and affordable for herders from the beginning and along their transhumant routes and include diffusion and rules within the regional regulatory framework about the need for herd treatments.