Uterus transplantation in the baboon: methodology and long-term function after auto-transplantation.

BACKGROUND: Techniques for uterus transplantation (UTx) have been developed in rodent/domestic animals towards future clinical introduction of UTx to treat uterine factor infertility. The aim of this study was to extend the UTx research into a non-human primate species by developing surgical techniques for uterus retrieval and transplantation in the baboon. METHODS: Female baboons (n = 15) underwent surgery, with the initial five animals used for studies of pelvic vascular anatomy. Retrieval surgery included isolation of the ovarian veins and the uterine arteries together with the anterior branches of the internal iliacs. The utero-tubal-ovarian specimen was removed, flushed and kept ex vivo for 2 h when the two arterial ends and two venous ends were anastomosed side-to-side to construct one arterial and one venous end. These were, at auto-transplantation, anastomosed end-to-side to the external iliacs and the animals (n = 10) were evaluated concerning cyclicity and later by laparoscopy/laparotomy. RESULTS: The total duration of organ retrieval, backtable preparation and transplantation was around 6 h with an overall ischaemic time of the specimen of about 3 h. One animal died due to cardiomyopathy. Five out of the nine surviving animals resumed cyclicity, as a sign of re-established ovarian function. Only two out of these five animals exhibited resumed menstruation, indicating re-established ovarian and uterine function. Laparoscopy confirmed normal-sized uteri in these two animals. CONCLUSIONS: This study demonstrates the feasibility of UTx by vascular anastomosis in a non-human primate species. The low success rate demonstrates the complexity involved in UTx surgery and the need for further methodological developments.

The detection of enteric viruses in selected urban and rural river water and sewage in Kenya, with special reference to rotaviruses.

AIM: To determine the occurrence of eight human enteric viruses in surface water and sewage samples from different geographical areas in Kenya. METHODS AND RESULTS: Enteric viruses were recovered from the water and sewage sources by glass-wool adsorption elution and/or polyethylene glycol/NaCl precipitation and detected by singleplex real-time and conventional PCR and reverse transcriptase-PCR assays. One or more enteric viruses were detected in nearly all sewage and river water samples except the urban Mbagathi River. The VP7 (G types) and the VP4 (P types) of the rotaviruses (RV) were characterized by multiplex nested PCR methods. The G and P types could be determined in 95·5% of the RV strains, respectively. Mixed G types were detected with G12 and G1 predominating, and unusual G types, G5 and G10, were present. P[4] predominated in the urban Karen sewage samples, while P[8] predominated in the urban and rural streams. CONCLUSIONS: The high prevalence of RVs in surface water highlights the importance of assessing the water sources used for domestic purposes for viral contamination. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the benefit of environmental surveillance as an additional tool to determine the epidemiology of RVs and other enteric viruses circulating in a given community.

Baboon serum is superior to human or bovine serum albumin for baboon sperm capacitation and zona binding.

BACKGROUND: Baboon in vitro fertilization requires capacitated sperm inappropriate media. In this study, we compared the effect of baboon serum (Bas), human serum albumin (HSA) and bovine serum albumin (BSA) on baboon sperm capacitation. METHODS: Five males (n = 5) were electroejaculated and 43 oocytes retrieved from super-ovulated female baboons (n = 10). Each sperm sample was assessed for initial motility and concentration before and after swim-up. For swim-up, each sperm sample was incubated separately in Biggers-Whitten-Whittingham media containing either BaS, HSA, BSA or without protein supplementation (control). After swim-up, each sperm aliquot was incubated with two to three oocytes. The number of sperm bound to the zona was evaluated after overnight incubation. RESULTS: Sperm motility and zona binding was significantly higher after capacitation in media supplemented with BaS than in HSA or BSA or in media without protein supplementation (P < 0.05). CONCLUSION: Baboon serum is superior to HSA or BSA for baboon sperm capacitation and zona binding.

Vaginal histological changes of the baboon during the normal menstrual cycle and pregnancy.

BACKGROUND: A baboon, a non-human primate, is phylogenetically close to human and has been used to study in detail aspects of reproductive physiology that cannot be studied in humans for ethical reasons. OBJECTIVE: To determine the histological changes in baboon vagina associated with cyclic variations during normal menstrual cycle. SETTING: The experiments were carried out at Institute of Primate Research (IPR), Karen, Nairobi, Kenya. SUBJECTS: Nine adult healthy female olive baboons were used in this study. These baboons were monitored over a period of one year and found to have regular menstrual cycles. The vaginal biopsies were taken at different menstrual stages, fixed in 10% formalin and processed to evaluate histological changes. RESULTS: Observation of the histological sections of the biopsies by light microscopy showed that there were histological changes associated with cyclic variations in female olive baboon. During the luteal phase, menstrual phase and pregnancy the squamous epithelium was very thin. The layer gradually thickens throughout the proliferative phase and was thickest during the ovulation period. CONCLUSION: The changes in squamous epithelium suggest that the baboon vagina undergoes histological changes throughout the menstrual cycle which may be associated with hormonal variations. The data from this study also suggest that olive baboon is a good model for investigating possible effects of hormonal contraceptives on vaginal epithelium and the mechanism of female heterosexual transmission of HIV.

Molecular characterisation of the rotavirus strains prevalent in Maua, Meru North, Kenya.

BACKGROUND: Rotavirus is the most common cause of severe infantile diarrhoea disease in infants and young children below five years worldwide. Rotavirus is associated with high cases of morbidity and mortality and it is estimated that up to 650,000 deaths in young children occur annually in the less developed countries and approximately 150,000-200,000 deaths occur in Africa alone. OBJECTIVE: To characterise the circulating rotavirus strains in Maua, Meru North district, Kenya. DESIGN: A prospective study to investigate and characterise rotavirus serotypes/genotypes and electropherotypes in infants and children with severe diarrhoea hospitalised and/or attending the outpatient department of Maua Methodist Hospital during the period April 2004 to September 2005. SETTING: Maua Methodist Hospital, Meru North, Kenya. SUBJECTS: Faecal samples were collected from 135 infants and children with acute diarrhoea and were screened first for the presence of human Group A rotavirus antigen using commercially available enzyme linked immunosorbent assay kit (ELISA). The positive samples were evaluated by sodium dodecyl polycrylamide gel electrophoresis (SDS-PAGE) to determine the viral RNA electropherotype profile. Rotavirus strains were also genotyped using reverse transcriptase polymerase chain reaction (RT-PCR) of the VP7 gene. RESULTS: Assay of these samples with commercial ELISA showed that 17.8% (24/135) were positive for group A rotavirus antigen. Twenty of these ELISA positive samples were also analysed by SDS-PAGE of which 75% (15/20) gave detectable electropherotype pattern with the long electropherotype being predominant 80.0% (12/15) followed by the short RNA profile 20.0% (2/ 15). Seventeen of the ELISA positive samples were genotyped for VP7 and the results showed that G9 was the most predominant genotype comprising 47.1% (8/17) followed by G8 29.4% (5/17), GI 17.4% (3/17) and the mixed genotype was G8/G9 5.9% (1/17). Most patients with rotavirus infection were of the age of 3 - 60 months, with 79% being less than 18 months old. CONCLUSION: The overall prevalence of rotavirus infection in young children with diarrhoea hospitalised and/or attending the out-patient department of Maua Methodist Hospital was 17.8% with the predominant serotype being G9. These results show that rotavirus plays an important role in severe viral diarrhoea in young children in Maua Meru North district, Kenya. Furthermore, this high G9 rotavirus prevalence in Kenya may require vaccine trials to be held in Kenya so as to determine the efficacy of new rotavirus vaccine candidates that do not include the G9 serotype.

Evidence for expression of endogenous retroviral sequences on primate reproductive tissues and detection of cross-reactive ERVS antigens in the baboon ovary: a review.

OBJECTIVE: To review recent research findings on the specific expression of endogenous retroviral sequences (ERVS) in reproductive tissues and their possible physiological roles. ERVS have been implicated in several biological events such as induction of resistance to exogenous retrovirus invasion, involvement in placental trophoblast formation, sperm maturation and differentiation; and stimulation of local immunosuppression to protect the foetus from immunological attack. DATA SOURCES: Critical review of relevant articles and abstracts cited in international and local journals, literature searches on Medline and Medchem up to 2005. DATA SYNTHESIS: Retroviruses have been implicated in the induction of tumour and immunological disorders. Over the years, endogenous retroviruses (ERVs) and retroviral elements have been detected in the genome of many vertebrate species, including primates. The evidence for the presence of retroviruses in the primate tissues such as the placenta, ovary, breast, testis and epididymis has been documented using electron microscopic studies. Retrovirus-like particles were found budding from the basal membrane of syncytiotrophoblasts, as well as in tumour cell lines in embryonic carcinoma or teratocarcinomas. Apart from their pathological effects, recent evidence suggests that these ERVs may play useful roles in normal physiological events. RESULTS: Recent studies indicate the expression of endogenous retroviruses in the testis, epididymis, placenta and breast. However, limited data exist on the detection of ERVs in the ovary. Overall, the precise functions for ERVs in these tissues are not well understood. In the testis and epididymis, speculative functions may include among others spermatogenesis and/or sperm maturation (differentiation) whereas in placenta they are possibly associated with trophoblast fusion and locally induced immunosuppression to protect the foetus from immunological attack. Experiments in our laboratory have indicated restricted expression of retroviral antigens including baboon endogenous retroviral proteins (BERV), ERV-3, HIV-1 gp41 and HERV-K env in the baboon ovary. CONCLUSION: ERVs are specifically expressed in different mammalian reproductive tissues and may have unique physiological roles.

Evidence for expression of endogenous retroviral sequences on primate reproductive tissues and detection of cross-reactive ervs antigens in the baboon ovary: a review

Objective: To review recent research findings on the specific expression of endogenous retroviral sequences (ERVS) in reproductive tissues and their possible physiological roles. ERVS have been implicated in several biological events such as induction of resistance to exogenous retrovirus invasion; involvement in placental trophoblast formation; sperm maturation and differentiation; and stimulation of local immunosuppression to protect the foetus from immunological attack. Data sources: Critical review of relevant articles and abstracts cited in international and local journals; literature searches on Medline and Medchem up to 2005. Data synthesis: Retroviruses have been implicated in the induction of tumour and immunological disorders. Over the years; endogenous retroviruses (ERVs) and retroviral elements have been detected in the genome of many vertebrate species; including primates. The evidence for the presence of retroviruses in the primate tissues such as the placenta; ovary; breast; testis and epididymis has been documented using electron microscopic studies. Retrovirus-like particles were found budding from the basal membrane of lyncytiotrophoblasts; as well as in tumour cell lines in embryonic carcinoma or teratocarcinomas. Apart from their pathological effects; recent evidence suggests that these ERVs may play useful roles in normal physiological events. Results: Recent studies indicate the expression of endogenous retroviruses in the testis; epididymis; placenta and breast. However; limited data exist on the detection of ERVs in the ovary. Overall; the precise functions for ERVs in these tissues are not well understood. In the testis and epididymis; speculative functions may include among others spermatogenesis and/or sperm maturation (differentiation) whereas in placenta they are possibly associated with trophoblast fusion and locally induced immunosuppression to protect the foetus from immunological attack. Experiments in our laboratory have indicated restricted expression of retroviral antigens including baboon endogenous retroviral proteins (BERV); ERV-3; HIV-1 gp41 and HERV-K env in the baboon ovary. Conclusion: ERVs are specifically expressed in different mammalian reproductive tissues and may have unique physiological roles

Serological detection of adenoviruses in non-human primates maintained in a colony in Kenya.

BACKGROUND: Adenoviruses are known to cause several human diseases including acute febrile respiratory syndromes, epidemic conjunctivitis and gastroenteritis. These diseases associated with adenovirus infection affect adults and are usually more severe in infants and children. Forty-seven human adenoviruses serotypes have so far been identified adenovirus. The diversity of these viruses has delayed progress on vaccine development due to difficulties in identifying appropriate vaccine targets. To date, limited studies have been done to determine the prevalence of adenovirus infection in non-human primates with the goal of developing a non-human primate model that can be used to study the mechanisms of infection. OBJECTIVE: To determine the prevalence of enteric adenovirus infection in Kenyan non-human primates. DESIGN: A prospective study to investigate the prevalence of enteric andenovirus infection in captive non-human primates maintained in a colony. SETTING: Faecal samples were collected from monkeys trapped from different geographical areas of Kenya and also from the ones maintained in a colony at the Institute of Primate Research (IPR), Kenya. SUBJECTS: Ninety four faecal samples were collected from three species of non-human primates consisting of various ages and sex. Samples were collected from monkeys trapped from different geographical areas of Kenya and also from the ones maintained in a colony at the Institute of Primate Research (IPR), Kenya. All the faecal samples were screened for presence of adenoviruses using a commercial antigen-capture enzyme immunoassay (EIA) kit, this is an enzyme-linked immunosorbent assay (ELISA) kit designed for diagnosis of human enteric adenoviruses in stool samples. RESULTS: The highest prevalence of adenoviruses, detected by EIA kit, was in olive baboons (Papio anubis, 52.9%), followed by vervet monkeys (Cercopithecus aethiops, 48.9%) and the yellow baboons (Papio cynocephalus, 18.8%). Sub-grouping within each species (based on age and sex) indicated no significant differences (p > 0.05) in adenovirus infection signifying equal susceptibility. The prevalence of adenoviruses in vervet monkeys that were also Simian Immunodeficiency virus (SIV) seropositive was determined and shown to be 63.2%. CONCLUSION: The results of this study indicate that adenovirus infection is prevalent among non-human primates in Kenya. These findings suggest that cross species transmission in Kenyan non-human primates may be a common occurrence and there is a possibility of zoonotic transmission of adenoviruses. Furthermore, our results highlight the potential of using these non-human primates as models for testing safety and efficacy of candidate adenovirus vaccines prior to clinical trials in humans.