Epidemiology and Molecular Characterization of Cryptosporidium spp. in Humans,Wild Primates,and Domesticated Animals in the Greater Gombe Ecosystem,Tanzania

Cryptosporidium is an important zoonotic parasite globally. Few studies have examined the ecology and epidemiology of this pathogen in rural tropical systems characterized by high rates of overlap among humans, domesticated animals, and wildlife. We investigated risk factors for Cryptosporidium infection and assessed cross-species transmission potential among people, non-human primates, and domestic animals in the Gombe Ecosystem, Kigoma District, Tanzania. A cross-sectional survey was designed to determine the occurrence and risk factors for Cryptosporidium infection in humans, domestic animals and wildlife living in and around Gombe National Park. Diagnostic PCR revealed Cryptosporidium infection rates of 4.3% in humans, 16.0% in non-human primates, and 9.6% in livestock. Local streams sampled were negative. DNA sequencing uncovered a complex epidemiology for Cryptosporidium in this system, with humans, baboons and a subset of chimpanzees infected with C. hominis subtype IfA12G2; another subset of chimpanzees infected with C. suis; and all positive goats and sheep infected with C. xiaoi. For humans, residence location was associated with increased risk of infection in Mwamgongo village compared to one camp (Kasekela), and there was an increased odds for infection when living in a household with another positive person. Fecal consistency and other gastrointestinal signs did not predict Cryptosporidium infection. Despite a high degree of habitat overlap between village people and livestock, our results suggest that there are distinct Cryptosporidium transmission dynamics for humans and livestock in this system. The dominance of C. hominis subtype IfA12G2 among humans and non-human primates suggest cross-species transmission. Interestingly, a subset of chimpanzees was infected with C. suis. We hypothesize that there is cross-species transmission from bush pigs (Potaochoerus larvatus) to chimpanzees in Gombe forest, since domesticated pigs are regionally absent. Our findings demonstrate a complex nature of Cryptosporidium in sympatric primates, including humans, and stress the need for further studies.     

Diet and niche breadth variation in the marbled parrotfish,Leptoscarus vaigiensis,among coral reef sites in Kenya

Studies on feeding ecology of fishes are important for understanding ecosystem structure and function. This study tested the hypothesis of diet and niche breath variation in the marbled parrotfish (Leptoscarus vaigiensis) among coral reefs of different protection levels in Kenya. Fish samples were obtained from protected (Malindi and Watamu marine parks), moderately fished (Malindi and Watamu marine reserves) and highly fished (Vipingo and Kanamai) reefs. Total lengths of fish samples were measured and their stomach contents quantified using the point method. Seasonal dietary composition, niche breaths and feeding intensities were compared between the sites using multivariate statistics. Results showed the parrotfish is a predominantly reef macroalgal grazer. Fish from protected sites fed on diverse dietary items compared to those from reserves and highly fished sites. Fish niche breadths differed between sites and seasons. Higher niche breadths occurred in protected sites during the north‐east monsoon, while higher values occurred at fished sites during the south‐east monsoon season. This study, the first of its kind in Kenya and most of the western Indian Ocean, describes feeding in the marbled parrotfish and spatial variation in niche breadth as influenced by fishing pressure, environmental variability and biological interactions.     

Microbial carbon oxidation rates and pathways in sediments of two Tanzanian mangrove forests

Temporal and spatial variations in benthic metabolism and anaerobic carbon oxidation pathways were assessed in an anthropogenically impacted (Mtoni) and a pristine (Ras Dege) mangrove forest in Tanzania. The objectives were (1) to evaluate how benthic metabolism is affected by organic carbon availability; (2) to determine the validity of diffusive release of CO₂ as a measure benthic carbon oxidation; and (3) to assess the partitioning of anaerobic carbon pathways and factors controlling the availability of electron acceptors (e.g. oxidized iron). Microbial carbon oxidation measured as diffusive exchange of O₂ and CO₂ (32?C67 and 28?C115 mmol m^?2 day^?1, respectively) showed no specific temporal patterns. Low intertidal sediments at Mtoni fed by labile algal carbon of anthropogenic origin had higher diffusive CO₂ release than high intertidal sediments that primarily received less reactive mangrove detritus. Diffusive release of CO₂ apparently underestimated total sediment carbon oxidation due to CO₂ loss from deep sediments via emission through biogenic structures (i.e. crab burrows and pneumatophores) and porewater seepage into creeks. We propose that diffusive fluxes in the present mangrove sediments are roughly equivalent to depth-integrated reactions occurring in the upper 12 cm. Anaerobic carbon oxidation was dominated by FeR irrespective of anthropogenic influence in sediments where the oxidizing effects of biogenic structures increased the Fe(III) level. More than 80% of the anaerobic carbon oxidation in Mtoni and Ras Dege sediments was due to FeR when reactive Fe(III) exceeded 30 ??mol cm^?3. The anthropogenic influence at Mtoni was primarily noted as up to one order of magnitude higher denitrification than at Ras Dege, but this process always accounted for less than 1% of total carbon oxidation. It is noteworthy that organic and nutrient enrichment of anthropogenic origin in Mtoni has no measurable effect on microbial processes, other than carbon oxidation in the low intertidal area and denitrification throughout the forest, and indicates a strong resilience of mangrove environments towards disturbances.     

Foci of endemic simian immunodeficiency virus infection in wild-living eastern chimpanzees (Pan troglodytes schweinfurthii)

Simian immunodeficiency virus of chimpanzees (SIVcpz) is the immediate precursor to human immunodeficiency virus type 1 (HIV-1), yet remarkably, the distribution and prevalence of SIVcpz in wild ape populations are unknown. Studies of SIVcpz infection rates in wild chimpanzees are complicated by the species' endangered status and by its geographic location in remote areas of sub-Saharan Africa. We have developed sensitive and specific urine and fecal tests for SIVcpz antibody and virion RNA (vRNA) detection and describe herein the first comprehensive prevalence study of SIVcpz infection in five wild Pan troglodytes schweinfurthii communities in east Africa. In Kibale National Park in Uganda, 31 (of 52) members of the Kanyawara community and 39 (of approximately 145) members of the Ngogo community were studied; none were found to be positive for SIVcpz infection. In Gombe National Park in Tanzania, 15 (of 20) members of the Mitumba community, 51 (of 55) members of the Kasekela community, and at least 10 (of approximately 20) members of the Kalande community were studied. Seven individuals were SIVcpz antibody and/or vRNA positive, and two others had indeterminate antibody results. Based on assay sensitivities and the numbers and types of specimens analyzed, we estimated the prevalence of SIVcpz infection to be 17% in Mitumba (95% confidence interval, 10 to 40%), 5% in Kasekela (95% confidence interval, 4 to 7%), and 30% in Kalande (95% confidence interval, 15 to 60%). For Gombe as a whole, the SIVcpz prevalence was estimated to be 13% (95% confidence interval, 7 to 25%). SIVcpz infection was confirmed in five chimpanzees by PCR amplification of partial pol and gp41/nef sequences which revealed a diverse group of viruses that formed a monophyletic lineage within the SIVcpzPts radiation. Although none of the 70 Kibale chimpanzees tested SIVcpz positive, we estimated the likelihood that a 10% or higher prevalence existed but went undetected because of sampling and assay limitations; this possibility was ruled out with 95% certainty. These results indicate that SIVcpz is unevenly distributed among P. t. schweinfurthii in east Africa, with foci or "hot spots" of SIVcpz endemicity in some communities and rare or absent infection in others. This situation contrasts with that for smaller monkey species, in which infection rates by related SIVs are generally much higher and more uniform among different groups and populations. The basis for the wide variability in SIVcpz infection rates in east African apes and the important question of SIVcpz prevalence in west central African chimpanzees (Pan troglodytes troglodytes) remain to be elucidated.     

The ranging patterns of elephants in Marsabit protected area,Kenya: the use of satellite‐linked GPS collars

We investigated the ranging patterns of elephants in the Marsabit protected area, north eastern Kenya, to ascertain the range of bachelor and female family herds in different seasons, and to identify corridor and noncorridor areas. Data were acquired for five bachelor and four female family herds equipped with satellite‐linked geographical positioning system collars, and monitored from December 2005 to December 2007. Distinct dry (about 260 km²) and wet seasons (about 910 km²) ranges were observed, with connecting corridors (north‐eastern corridor: about 90 km long, about 2‐7 km wide; southern corridors: about 10‐20 km long, about 2‐3 km wide). The dry season range corresponded with Marsabit evergreen forest, while the wet season range matched with dry deciduous lowland shrubs. The ranging elephants moved at speed of about 0.2‐20 kmh^?1. Bachelor herds moved faster than female family herds. Elephants moved fast during the intermediate and wet seasons than during the dry season. The speed of ranging elephants was over 1 kmh^?1 in the corridor areas and about 0.2 to less than 1 kmh^?1 in the non‐corridor areas. Expansion of settlements towards corridor areas needs to be controlled to avoid future blocking of connectivity between wet and dry season elephant ranges.     

Ferrocene tripeptide Gly-Pro-Arg conjugates: Synthesis and inhibitory effects on Alzheimer’s Aβ1–42 fibrillogenesis and Aβ-induced cytotoxicity in vitro

Alzheimer’s disease (AD) is the most common cause of dementia, and currently there is no clinical treatment to cure it or to halt its progression. Aggregation and fibril formation of β-amyloid peptides (Aβ) are central events in the pathogenesis of AD. Many efforts have been spent on the development of effective inhibitors to prevent Aβ fibrillogenesis and cause disaggregation of preformed Aβ fibrils. In this study, the conjugates of ferrocene and Gly-Pro-Arg (GPR) tripeptide, Boc-Gly-Pro-Arg(NO₂)-Fca-OMe (4, GPR–Fca) and Fc-Gly-Pro-Arg-OMe (7, Fc–GPR) (Fc: ferrocene; Fca: ferrocene amino acid) were synthesized by HOBT/HBTU protocol in solution. These ferrocene GPR conjugates were employed to inhibit Aβ_1–42 fibrillogenesis and to disaggregate preformed Aβ fibrils. The inhibitory properties of ferrocene GPR conjugates on Aβ_1–42 fibrillogenesis were evaluated by thioflavin T (ThT) fluorescence assay, and confirmed by atomic force microscopy (AFM) analysis. The interaction between the ferrocene GPR conjugates and Aβ_1–42 was monitored by electrochemical means. Our results showed that both GPR and GPR–Fca can significantly inhibit the fibril formation of Aβ_1–42, and cause disaggregation of the preformed fibrils. As expected, GPR–Fca shows stronger inhibitory effect on Aβ_1–42 fibrillogenesis than that of its parent peptide GPR. In contrast, Fc–GPR shows no inhibitory effect on fibrillogenesis of Aβ_1–42. Furthermore, GPR–Fca demonstrates significantly protection against Aβ-induced cytotoxicity and exhibits high resistance to proteolysis and good lipophilicity.     

Global Positioning System Data-Loggers: A Tool to Quantify Fine-Scale Movement of Domestic Animals to Evaluate Potential for Zoonotic Transmission to an Endangered Wildlife Population

Domesticated animals are an important source of pathogens to endangered wildlife populations, especially when anthropogenic activities increase their overlap with humans and wildlife. Recent work in Tanzania reports the introduction of Cryptosporidium into wild chimpanzee populations and the increased risk of ape mortality associated with SIVcpz-Cryptosporidium co-infection. Here we describe the application of novel GPS technology to track the mobility of domesticated animals (27 goats, 2 sheep and 8 dogs) with the goal of identifying potential routes for Cryptosporidium introduction into Gombe National Park. Only goats (5/27) and sheep (2/2) were positive for Cryptosporidium. Analysis of GPS tracks indicated that a crop field frequented by both chimpanzees and domesticated animals was a potential hotspot for Cryptosporidium transmission. This study demonstrates the applicability of GPS data-loggers in studies of fine-scale mobility of animals and suggests that domesticated animal–wildlife overlap should be considered beyond protected boundaries for long-term conservation strategies.     

Molecular ecology and natural history of simian foamy virus infection in wild-living chimpanzees

Identifying microbial pathogens with zoonotic potential in wild-living primates can be important to human health, as evidenced by human immunodeficiency viruses types 1 and 2 (HIV-1 and HIV-2) and Ebola virus. Simian foamy viruses (SFVs) are ancient retroviruses that infect Old and New World monkeys and apes. Although not known to cause disease, these viruses are of public health interest because they have the potential to infect humans and thus provide a more general indication of zoonotic exposure risks. Surprisingly, no information exists concerning the prevalence, geographic distribution, and genetic diversity of SFVs in wild-living monkeys and apes. Here, we report the first comprehensive survey of SFVcpz infection in free-ranging chimpanzees (Pan troglodytes) using newly developed, fecal-based assays. Chimpanzee fecal samples (n = 724) were collected at 25 field sites throughout equatorial Africa and tested for SFVcpz-specific antibodies (n = 706) or viral nucleic acids (n = 392). SFVcpz infection was documented at all field sites, with prevalence rates ranging from 44% to 100%. In two habituated communities, adult chimpanzees had significantly higher SFVcpz infection rates than infants and juveniles, indicating predominantly horizontal rather than vertical transmission routes. Some chimpanzees were co-infected with simian immunodeficiency virus (SIVcpz); however, there was no evidence that SFVcpz and SIVcpz were epidemiologically linked. SFVcpz nucleic acids were recovered from 177 fecal samples, all of which contained SFVcpz RNA and not DNA. Phylogenetic analysis of partial gag (616 bp), pol-RT (717 bp), and pol-IN (425 bp) sequences identified a diverse group of viruses, which could be subdivided into four distinct SFVcpz lineages according to their chimpanzee subspecies of origin. Within these lineages, there was evidence of frequent superinfection and viral recombination. One chimpanzee was infected by a foamy virus from a Cercopithecus monkey species, indicating cross-species transmission of SFVs in the wild. These data indicate that SFVcpz (i) is widely distributed among all chimpanzee subspecies; (ii) is shed in fecal samples as viral RNA; (iii) is transmitted predominantly by horizontal routes; (iv) is prone to superinfection and recombination; (v) has co-evolved with its natural host; and (vi) represents a sensitive marker of population structure that may be useful for chimpanzee taxonomy and conservation strategies.     

Two jacalin-related lectins from seeds of the African breadfruit (Treculia africana L.)

Two jacalin-related lectins (JRLs) were purified by mannose-agarose and melibiose-agarose from seeds of Treculia africana. One is galactose-recognizing JRL (gJRL), named T. africana agglutinin-G (TAA-G), and another one is mannose-recognizing JRL (mJRL), TAA-M. The yields of the two lectins from the seed flour were approximately 7.0 mg/g for gJRL and 7.2 mg/g for mJRL. The primary structure of TAA-G was determined by protein sequencing of lysyl endopeptic peptides and chymotryptic peptides. The sequence identity of TAA-G to other gJRLs was around 70%. Two-residue insertion was found around the sugar-binding sites, compared with the sequences of other gJRLs. Crystallographic studies on other gJRLs have shown that the primary sugar-binding site of gJRLs can accommodate Gal, GalNAc, and GalNAc residue of T-antigen (Galβ1-3GalNAcα-). However, hemagglutination inhibition and glycan array showed that TAA-G did not recognize GalNAc itself and T-antigen. TAA-G preferred melibiose and core 3 O-glycan.     

Comparative Metabolomic Profiling Reveals That Dysregulated Glycolysis Stemming from Lack of Salvage NAD+ Biosynthesis Impairs Reproductive Development in Caenorhabditis elegans

Temporal developmental progression is highly coordinated in Caenorhabditis elegans. However, loss of nicotinamidase PNC-1 activity slows reproductive development, uncoupling it from its typical progression relative to the soma. Using LC/MS we demonstrate that pnc-1 mutants do not salvage the nicotinamide released by NAD⁺ consumers to resynthesize NAD⁺, resulting in a reduction in global NAD⁺ bioavailability. We manipulate NAD⁺ levels to demonstrate that a minor deficit in NAD⁺ availability is incompatible with a normal pace of gonad development. The NAD⁺ deficit compromises NAD⁺ consumer activity, but we surprisingly found no functional link between consumer activity and reproductive development. As a result we turned to a comparative metabolomics approach to identify the cause of the developmental phenotype. We reveal widespread metabolic perturbations, and using complementary pharmacological and genetic approaches, we demonstrate that a glycolytic block accounts for the slow pace of reproductive development. Interestingly, mitochondria are protected from both the deficiency in NAD⁺ biosynthesis and the effects of reduced glycolytic output. We suggest that compensatory metabolic processes that maintain mitochondrial activity in the absence of efficient glycolysis are incompatible with the requirements for reproductive development, which requires high levels of cell division. In addition to demonstrating metabolic requirements for reproductive development, this work also has implications for understanding the mechanisms behind therapeutic interventions that target NAD⁺ salvage biosynthesis for the purposes of inhibiting tumor growth.