Assessing maternal and neonatal near-miss reviews in rural Nepal: an implementation research study to inform scale-up.

AIM: To understand how maternal and neonatal near-miss reviews could be implemented and scaled-up in rural communities through the existing district health system in Nepal. METHODS: Mixed methods with a modified time series evaluation design were used. The World Health Organization maternal and neonatal near-miss criteria used in multicountry surveys were adapted and used to define maternal and neonatal near-miss cases. RESULTS: The World Health Organization near-miss criteria were mainly applicable at the district hospital setting, but further adaptations were needed for community-level birthing centres, as organ dysfunction and critical intervention criteria were not found appropriate. In birthing centres, disease-based criteria were applicable for maternal near-miss review, and danger and clinical sign-based and condition at birth criteria were applicable for neonatal near-miss review. Primary barriers to implementation were attrition of trained staff due to the frequent transfer of healthcare providers, and time constraints of district hospital medical doctors for case-by-case reviews as they were often busy in hospital and in their private clinics. CONCLUSION: Adapted maternal and neonatal near-miss review process implementation in Nepal is feasible through the existing government health system.

Intraspecific and interspecific genetic variation of Gongylonema pulchrum and two rodent Gongylonema spp. (G. aegypti and G. neoplasticum), with the proposal of G. nepalensis n. sp. for the isolate in water buffaloes from Nepal.

The gullet worm (Gongylonema pulchrum) has been recorded from a variety of mammals worldwide. In an earlier study, we demonstrated two separate transmission cycles in cattle (Bos taurus) and wild mammals in Japan based on nucleotide sequences of the ribosomal RNA gene (rDNA) and cytochrome c oxidase subunit I (cox-1) region of mitochondrial DNA of multiple isolates of different origins. Our earlier study additionally demonstrated two major cox-1 haplotypes of G. pulchrum prevalent in cattle in Japan. In the present study, we collected G. pulchrum from cattle and goats (Capra hircus) in Alashan League, Inner Mongolia, China; Gongylonema aegypti from spiny mice (Acomys dimidiatus) in the Sinai Peninsula, Egypt; and Gongylonema neoplasticum from a black rat (Rattus rattus) in Okinawa Island, Japan, to analyze their genetic relationships with G. pulchrum in Japan. The gullet worms from Alashan League had almost identical rDNA nucleotide sequences and two cox-1 haplotypes as seen in G. pulchrum from the cattle in Japan. The two rodent Gongylonema spp. had distinct rDNA nucleotide sequences compared with those of G. pulchrum; only the 18S and 5.8S rDNA sequences showed high identities at 97.2-98.7%, while the remaining sequences were less than 75% identical. The 18S, 5.8S, and 28S rDNA sequences of the two rodent Gongylonema spp. showed nucleotide identities of 99.8% (1811/1814), 100% (158/158), and 98.9% (3550/3590), respectively. The cox-1 regions showed 91.6% (338/369)-92.1% (340/369) identities, with completely identical amino acid sequences. The genetic diversities of three distinct Gongylonema spp. and their possible intraspecific genetic variation may allow us to resolve the taxonomic position of Gongylonema spp. which display few obvious morphological differences from their congeners. Consequently, the Gongylonema isolate from water buffaloes (Bubalus bubalis) in Nepal reported in our previous study is concluded to be a new species, and Gongylonema nepalensis n. sp. is erected for it.

A distinct genetic population of Gongylonema pulchrum from water buffaloes in Nepal.

Whole-length esophagi of 111 Murrah cross water buffaloes (Bubalus bubalis) were collected in the Kathmandu and Chitwan districts of Nepal from December 2009 to February 2010. Gullet worms showing a typical epithelium-dwelling character were detected in 13 of 53 (24.5%) buffaloes in Kathmandu and in 5 of 58 (8.6%) buffaloes in Chitwan. The worms' morphology and measurements were identical to those of Gongylonema pulchrum Molin, 1857, except for the length of the left spicules relative to the body length. Scanning electron microscopy did not detect any further morphological differences regarding the collected specimen from Nepal compared with G. pulchrum . The ribosomal RNA gene (rDNA), including internal transcribed spacer (ITS) 1 and 2, and a partial region of the cytochrome c oxidase subunit I (COI) of mitochondrial DNA of the worms were characterized and compared with those of G. pulchrum collected from cattle, deer, wild boars, and monkeys in Japan and from cattle in Iran. The 18S, 5.8S, and 28S rDNA nucleotide sequences of the buffalo-collected worms had 99.8% (1,779/1,782), 100% (158/158), and 98.3-98.8% (3,494-3,507/3,551) identities, respectively, with those of G. pulchrum from the other host mammals. The ITS regions exhibited higher variations between the buffalo-collected worms and G. pulchrum from the other host mammals (85-88% identity for ITS1 and 56-80% identity for ITS2). The COI also showed lower identities (89.2-90.2%), although only a single amino acid substitution was noted compared with the majority of G. pulchrum samples collected in Japan. Based on these molecular genetic characters in the rDNA and COI mitochondrial DNA, together with a shorter left spicule length relative to body length, the gullet worms isolated from buffaloes in Nepal might belong to a distinct local or buffalo-preferring population of G. pulchrum, although its geographical distribution on the continent and host specificity remain to be clarified.