Christopher A. Whittier

Molecular and morphologic characterization of a Cryptosporidium genotype identified in lemurs

This study reports the molecular and morphologic characterization of a Cryptosporidium sp., identified in stools of captive lemurs Propithecus verreauxi coquereli. Stool samples were collected from seven animals (n=7) presenting episodes of diarrhea. Bright-field light microscopy of stool smears stained with modified acid-fast technique revealed the presence of Cryptosporidium sp. oocysts in four of the stool samples analyzed. All microscopically positive samples were confirmed by PCR using primers designed to amplify DNA fragments from two independent loci, i.e. the Cryptosporidium oocyst wall protein (COWP) gene and the small subunit ribosomal RNA (ssrRNA) gene. Phylogenetic analysis based on the full-length ssrRNA gene placed this isolate within a clade that contains all currently known C. parvum species/genotypes, closely related to the C. parvum pig genotype. Comparison with partial ssrRNA sequences available in the GenBank revealed 100% sequence identity with the genotype previously identified in Canadian patients. This finding was confirmed further by comparison of the COWP gene partial sequences.

Comparison of DNA extraction methods for PCR amplification of mitochondrial cytochrome c oxidase subunit II (COII) DNA from primate fecal samples

Mitochondrial COII DNA was amplified by PCR from total DNA extracted from field collected primate fecal samples (n=24) which had been stored without refrigeration for over 30 days. High molecular weight DNA total DNA was obtained from samples stored in 70% (v/v) ethanol, SDS lysis buffer (LB) and guanidine isothiocyanate buffer (GTB) than from samples stored in 10% formalin. Fecal DNA quality and COII amplification varied according to storage solution (formalin, ethanol, LB and GTB), extraction method (LB-based and GTB-based) and primate species (chimpanzee, baboon, human). It is recommended that fecal samples be collected in LB for DNA analysis. However, GTB-based protocols are suitable when total RNA is needed for epidemiological studies of viral diseases or gene expression analysis.

Proximate composition of milk from free-ranging mountain gorillas (Gorilla beringei beringei)

Published data on milk composition for nonhuman primates, especially great apes, are lacking. Milk composition data are important for understanding the physiology and evolution of mammalian milk production, as well as the nutritional requirements of infants. For many primate species these data have added relevance because of the need to hand raise infants orphaned by poaching or separated from their mothers in captivity. The proximate composition (dry matter (DM), protein, fat, sugar) of free-ranging mountain gorilla (MG) (Gorilla beringei beringei) milk was characterized from samples (N = 10) collected opportunistically during field procedures. The mean values for mid-lactation (1-50 months) milk samples from healthy MGs (N = 7) were: 10.7% DM, 1.9% fat, 1.4% crude protein, 6.8% sugar, and 0.53 kcal/g. These results are lower in fat and total energy than most other Hominidae, including humans. One early-lactation sample was high in protein content while the composition of two samples from gorillas with poor health and suspected poor milk quality both deviated from the normal, mid-lactation pattern. This survey adds to the data available for primate milk composition and suggests that wild MG infants normally consume milk that is lower in fat and total energy than human milk.

Occupational health and gorilla conservation in Rwanda.

Abstract The design and implementation of an employee health program for people who work with mountain. Gorillas in Rwanda is described. This program aims to improve worker health and to reduce human-to-gorilla transmission of infectious disease. The program covered approximately III workers, generally heal thy men and women 25–45 years old, including essentially all people in Rwanda who have regular contact with gorillas. Initial assessment included a questionnaire, medical examination, and local tests. U.S. laboratory facilities were utilized to confirm some results and for serologic testing for zoonotic (simian) viruses. Initial interventions included STD/HIV prevention health education, tetanus immunization, and anthelminthic treatment. Local physicians continue to provide health services, including follow-up testing and treatment. Mountain Gorilla Veterinary Project (MGVP) veterinarians assist in planning and implementing continuing program components in collaboration with local health authorities and the other employing organizations.

Effect of Antibiotic Treatment on the Gastrointestinal Microbiome of Free-Ranging Western Lowland Gorillas (Gorilla g. gorilla)

The mammalian gastrointestinal (GI) microbiome, which plays indispensable roles in host nutrition and health, is affected by numerous intrinsic and extrinsic factors. Among them, antibiotic (ATB) treatment is reported to have a significant effect on GI microbiome composition in humans and other animals. However, the impact of ATBs on the GI microbiome of free-ranging or even captive great apes remains poorly characterized. Here, we investigated the effect of cephalosporin treatment (delivered by intramuscular dart injection during a serious respiratory outbreak) on the GI microbiome of a wild habituated group of western lowland gorillas (Gorilla gorilla gorilla) in the Dzanga Sangha Protected Areas, Central African Republic. We examined 36 fecal samples from eight individuals, including samples before and after ATB treatment, and characterized the GI microbiome composition using Illumina-MiSeq sequencing of the bacterial 16S rRNA gene. The GI microbial profiles of samples from the same individuals before and after ATB administration indicate that the ATB treatment impacts GI microbiome stability and the relative abundance of particular bacterial taxa within the colonic ecosystem of wild gorillas. We observed a statistically significant increase in Firmicutes and a decrease in Bacteroidetes levels after ATB treatment. We found disruption of the fibrolytic community linked with a decrease of Ruminoccocus levels as a result of ATB treatment. Nevertheless, the nature of the changes observed after ATB treatment differs among gorillas and thus is dependent on the individual host. This study has important implications for ecology, management, and conservation of wild primates.

Real-time PCR detection of Campylobacter spp. in free-ranging mountain gorillas (Gorilla beringei beringei).

Health monitoring of wildlife populations can greatly benefit from rapid, local, noninvasive molecular assays for pathogen detection. Fecal samples collected from free-living Virunga mountain gorillas (Gorilla beringei beringei) between August 2002 and February 2003 were tested for Campylobacter spp. DNA using a portable, real-time polymerase chain reaction (PCR) instrument. A high prevalence of Campylobacter spp. was detected in both individually identified (22/26=85%) and nest-collected samples (68/114=59.6%), with no statistically significant differences among different gorilla sexes or age classes or between tourist-visited versus research gorilla groups. The PCR instrument was able to discriminate two distinct groups of Campylobacter spp. in positive gorilla samples based on the PCR product fluorescent-probe melting profiles. The rare type (6/90 positives, 7%, including three mixed cases) matched DNA sequences of Campylobacter jejuni and was significantly associated with abnormally soft stools. The more common type of positive gorilla samples (87/90 positives, 97%) were normally formed and contained a Campylobacter sp. with DNA matching no published sequences. We speculate that the high prevalence of Campylobacter spp. detected in gorilla fecal samples in this survey mostly reflects previously uncharacterized and nonpathogenic intestinal flora. The real-time PCR assay was more sensitive than bacterial culture with Campylobacter-specific media and commercially available, enzyme immunoassay tests for detecting Campylobacter spp. in human samples.

Occupational Risks and Exposures Among Wildlife Health Professionals

Most emerging infectious diseases are zoonotic in origin, with wildlife a frequent source of zoonotic disease events. Although individuals with extensive wildlife contact may be at the greatest risk of contracting novel infectious agents, the occupational risk of those working closely with wildlife has not been well studied. This study assessed the occupational exposures among wildlife health professionals working in multiple countries worldwide. An occupational risk survey of past and present exposures was developed and administered online in a confidential manner to wildlife workers recruited through an ongoing international wildlife pathogen surveillance project. Surveys were completed by 71 participants in 14 countries. Significant lifetime exposures reported included bites from bats and rodents and touching dead animals. Completion of training in occupational safety was reported by 75% of respondents. While gloves were used for most tasks, use of N95 respirators and other personal protective equipment varied by task. Eighty percent of workers reported rabies vaccination. Some respondents indicated interest in enhanced occupational health services targeting their unique needs. Wildlife workers represent an occupational population at risk of zoonotic infection and injury. Enhanced occupational health services targeting wildlife workers could reduce the risk and sequelae of zoonotic exposure and infection.