Characterization of major histocompatibility complex class I, and class II DRB loci of captive and wild Indian leopards (Panthera pardus fusca).

The major histocompatibility complex (MHC), in vertebrate animals, is a multi-genic protein complex that encodes various receptors. During a disease, MHC interacts with the antigen and triggers a cascade of adaptive immune responses to overcome a disease outbreak. The MHC is very important region from immunological point of view, but it is poorly characterized among Indian leopards. During this investigation, we examined genetic diversity for MHC class I (MHC-I) and MHC class II-DRB (MHC-II) among wild and captive Indian leopards. This study estimated a pool of 9 and 17 alleles for MHC-I and MHC-II, respectively. The wild group of individuals showed higher nucleotide diversity and amino acid polymorphism compared to the captive group. A phylogenetic comparison with other felids revealed a clustering in MHC-I and interspersed presence in MHC-II sequences. A test for selection also revealed a deviation from neutrality at MHC-II DRB loci and higher non-synonymous substitution rate (dN) among the individuals from wild group. Further, the wild individuals showed higher dN for both MHC I and II genes compared to the group that was bred under captive conditions. These findings suggest the role of micro-evolutionary forces, such as pathogen-mediated selection, to cause MHC variations among the two groups of Indian leopards, because the two groups have been bred in two different environments for a substantial period of time. Since, MHC diversity is often linked with the quality of immunological health; the results obtained from this study fill the gap of knowledge on disease predisposition among wild and captive Indian leopards.

Range-Wide Snow Leopard Phylogeography Supports Three Subspecies.

The snow leopard, Panthera uncia, is an elusive high-altitude specialist that inhabits vast, inaccessible habitat across Asia. We conducted the first range-wide genetic assessment of snow leopards based on noninvasive scat surveys. Thirty-three microsatellites were genotyped and a total of 683 bp of mitochondrial DNA sequenced in 70 individuals. Snow leopards exhibited low genetic diversity at microsatellites (AN = 5.8, HO = 0.433, HE = 0.568), virtually no mtDNA variation, and underwent a bottleneck in the Holocene (∼8000 years ago) coinciding with increased temperatures, precipitation, and upward treeline shift in the Tibetan Plateau. Multiple analyses supported 3 primary genetic clusters: (1) Northern (the Altai region), (2) Central (core Himalaya and Tibetan Plateau), and (3) Western (Tian Shan, Pamir, trans-Himalaya regions). Accordingly, we recognize 3 subspecies, Panthera uncia irbis (Northern group), Panthera uncia uncia (Western group), and Panthera uncia uncioides (Central group) based upon genetic distinctness, low levels of admixture, unambiguous population assignment, and geographic separation. The patterns of variation were consistent with desert-basin "barrier effects" of the Gobi isolating the northern subspecies (Mongolia), and the trans-Himalaya dividing the central (Qinghai, Tibet, Bhutan, and Nepal) and western subspecies (India, Pakistan, Tajikistan, and Kyrgyzstan). Hierarchical Bayesian clustering analysis revealed additional subdivision into a minimum of 6 proposed management units: western Mongolia, southern Mongolia, Tian Shan, Pamir-Himalaya, Tibet-Himalaya, and Qinghai, with spatial autocorrelation suggesting potential connectivity by dispersing individuals up to ∼400 km. We provide a foundation for global conservation of snow leopard subspecies, and set the stage for in-depth landscape genetics and genomic studies.

Comparison of LED-Based Transillumination Device with Traditional Vein Viewing Methods for Difficult Intravenous Cannulation in Indian Children: A Nonrandomized Controlled Trial.

OBJECTIVES: To compare the utility of transillumination device with traditional vein viewing in situations with difficult peripheral venous access in pediatric patients. METHODS: This was a nonrandomized, controlled trial. All the children aged between 3 to 36 mo admitted in tertiary care referral hospital, who satisfied difficult intravenous access (DIVA) score of equal to or more than 4 were included in the study. The children were assigned to transillumination device group (intervention) and traditional vein viewing group (traditional). The proportion of successful cannulation in the first attempt and median number attempts required to successfully cannulate in each group were estimated. RESULTS: A total of 509 children were included in the study. The proportion of single attempt cannulation was significantly higher in the intervention group as compared to traditional group (p value = 0.001). The median number of attempts to successfully cannulate was found to be significantly less in the interventional group (median 1 vs. 2; p value = 0.001). On bivariate analysis, use of transillumination device was found to have a 2.64 times higher likelihood to successfully cannulate in the first attempt. CONCLUSION: The use of transillumination device significantly improves the first attempt success rate and number of attempts for successful cannulation.

Adverse events associated with Measles and Rubella vaccination campaign 2019 in India.

PURPOSE: The purpose of this study is to study the various adverse reactions caused post measles and rubella vaccination done during measles rubella (MR) vaccine campaign in India. MATERIALS AND METHODS: Prospective, observational study was done in a government tertiary care pediatric intensive care unit. Children aged between 9 months to 15 years, who presented with adverse effects (severe enough to warrant admission) within 7 days of MR vaccine administration. RESULTS: Most common presenting complaint was fever (44.8%), followed by vomiting (34.5%), abdominal pain and dizziness (31%). Abnormal body movements were noted in two children (6.8%) on first day and in one child on fifth day of vaccine administration. Two children (6.8%) presented with generalized macular rashes all over the body on 4th day after vaccination. Altered sensorium on same day of vaccine administration was the presenting symptom of one child. All children improved gradually and were discharged after few days with no mortality or long-term morbidity. Investigations were done according to the protocol of the unit; nothing came significant to be reported. Neither of the children had positive blood culture. CONCLUSION: MR vaccination programs are scientifically sound, highly recommended and proven effective globally. Causality assessment of adverse events is still an evolving science, and despite taking all the measures and adopting all the available scientific methods, sometimes it is not possible to incontrovertibly prove the causal association of an event with a vaccine. Much more advancement in this area is needed.

Ancient mtDNA from the extinct Indian cheetah supports unexpectedly deep divergence from African cheetahs.

The Indian cheetah was hunted to extinction by the mid-20th century. While analysis of 139 bp of mitochondrial DNA (mtDNA) has confirmed that the Indian cheetah was part of the Asiatic subspecies (Acinonyx jubatus venaticus), the detailed relationships between cheetah populations remains unclear due to limited genetic data. We clarify these relationships by studying larger fragments of cheetah mtDNA, both from an Indian cheetah museum specimen and two African cheetah, one modern and one historic, imported into India at different times. Our results suggest that the most recent common ancestor of cheetah mtDNA is approximately twice as ancient as currently recognised. The Indian and Southeast African (Acinonyx jubatus jubatus) cheetah mtDNA diverged approximately 72 kya, while the Southeast and Northeast African (Acinonyx jubatus soemmeringii) cheetah mtDNA diverged around 139 kya. Additionally, the historic African cheetah sampled from India proved to have an A. j. jubatus haplotype, suggesting a hitherto unrecognised South African route of cheetah importation into India in the 19th century. Together, our results provide a deeper understanding of the relationships between cheetah subspecies, and have important implications for the conservation of A. j. venaticus and potential reintroduction of cheetahs into India.

Characteristics of the complete mitochondrial genome of the monotypic genus Arctictis (Family: Viverridae) and its phylogenetic implications.

The binturong (Arctictis binturong) is classified as a member of the subfamily Paradoxurinae within the family Viverridae (Carnivora: Mammalia) and comprises nine subspecies spread across Southern and Southeast Asia. Here, we describe the complete mitochondrial genome of the Indian subspecies A. b. albifrons using next-generation sequencing methods. The total length of the A. b. albifrons mitogenome was 16,642 bp. Phylogenetic analyses based on 13 mitochondrial protein-coding genes placed the binturong as a sister taxon to Paguma larvata within the Paradoxurinae and supported the clustering of Genettinae and Viverrinae and the monophyly of Viverridae and six other families of feliforms, consistent with previous studies. Divergence time estimates suggest that the Viverridae diversified during the Miocene (22.62 Mya: 95% CI [20.78-24.54] Mya) and that Arctictis and Paguma split 12.57 Mya (95% CI [8.66-15.67] Mya). Further molecular studies are required to test the distinctiveness and diversity of the nine putative subspecies of binturong.

Indian Academy of Pediatrics Guidelines on the Fast and Junk Foods, Sugar Sweetened Beverages, Fruit Juices, and Energy Drinks.

JUSTIFICATION: In view of easy availability and increasing trend of consumption of fast foods and sugar sweetened beverages (fruit juices and drinks, carbonated drinks, energy drinks) in Indian children, and their association with increasing obesity and related non-communicable diseases, there is a need to develop guidelines related to consumption of foods and drinks that have the potential to increase this problem in children and adolescents. OBJECTIVES: To review the evidence and formulate consensus statements related to terminology, magnitude of problem and possible ill effects of junk foods, fast foods, sugar-sweetened beverages and carbonated drinks; and to formulate recommendations for limiting consumption of these foods and beverages in Indian children and adolescents. PROCESS: A National Consultative group constituted by the Nutrition Chapter of the Indian Academy of Pediatrics (IAP), consisting of various stakeholders in private and public sector, reviewed the literature and existing guidelines and policy regulations. Detailed review of literature was circulated to the members, and the Group met on 11th March 2019 at New Delhi for a day-long deliberation on framing the guidelines. The consensus statements and recommendations formulated by the Group were circulated to the participants and a consensus document was finalized. CONCLUSIONS: The Group suggests a new acronym 'JUNCS' foods, to cover a wide variety of concepts related to unhealthy foods (Junk foods, Ultra-processed foods, Nutritionally inappropriate foods, Caffeinated/colored/carbonated foods/beverages, and Sugar-sweetened beverages). The Group concludes that consumption of these foods and beverages is associated with higher free sugar and energy intake; and is associated with higher body mass index (and possibly with adverse cardiometabolic consequences) in children and adolescents. Intake of caffeinated drinks may be associated with cardiac and sleep disturbances. The Group recommends avoiding consumption of the JUNCS by all children and adolescents as far as possible and limit their consumption to not more than one serving per week. The Group recommends intake of regional and seasonal whole fruits over fruit juices in children and adolescents, and advises no fruit juices/drinks to infants and young children (age <2y), whereas for children aged 2-5 y and >5-18 y, their intake should be limited to 125 mL/day and 250mL/day, respectively. The Group recommends that caffeinated energy drinks should not be consumed by children and adolescents. The Group supports recommendations of ban on sale of JUNCS foods in school canteens and in near vicinity, and suggests efforts to ensure availability and affordability of healthy snacks and foods. The Group supports traffic light coding of food available in school canteens and recommends legal ban of screen/print/digital advertisements of all the JUNCS foods for channels/magazines/websites/social media catering to children and adolescents. The Group further suggests communication, marketing and policy/taxation strategies to promote consumption of healthy foods, and limit availability and consumption of the JUNCS foods.

The complete mitochondrial genome of Asian palm civet (Paradoxurus hermaphroditus) with phylogenetic consideration.

Asian palm civet (Paradoxurus hermaphroditus) is one of the smallest palm civet which is least studied. Here, we report the first complete mitochondrial (mt) genome of Asian palm civet (P. hermaphroditus). The circular mt genome with a length of 16,706 bp contained 1 control region, 2 rRNAs, 13 protein-coding genes, and 22 tRNAs. Overall base composition of the complete mt DNA was 33.7% A, 30.5% T, 22.9% C, and 12.9% G. All the genes in mt genome of Asian palm civet (P. hermaphroditus) were distributed on the H-strand, except ND6 and eight tRNA genes encoded on the L-strand. Maximum likelihood (ML) and Bayesian inference (BI) methods were used to infer the phylogenetic relationship of P. hermaphroditus. The phylogenetic analysis shows that all species from the family Viverridae cluster together, in which P. hermaphroditus exhibits the closest relationship with P. larvata.

Characterization of the complete mitogenome of Indian Mouse Deer, Moschiola indica (Artiodactyla: Tragulidae) and its evolutionary significance.

The mitochondrial genome of Indian mouse deer (Moschiola indica) was sequenced, assembled and characterized for the first time using 22 pairs of polymerase chain reaction (PCR) primers. The mitogenome of M. indica which is 16,444 bp in size was found very similar to most vertebrates in organisation that harbours 13 protein-coding genes, 22 transfer RNA, 2 ribosomal RNA and 1A + T-rich region. Its comparison with over 52 mitogenomes of the order Artiodactyla, showed a conserved nature of gene organisation, codon usage, gene orientation and evolutionary rates of proteins except that M. indica possesses an extra copy of trnF. The complete mitogenome and protein-coding genes of M. indica were found to be highly A + T biased. Rate of protein evolution was highest in atp8 and lowest in cox3. Further, a higher purifying selection pressure was found to be acting on family Tragulidae compared to Bovidae and Cervidae. The phylogenetic analysis of M. indica placed the Tragulidae as sister-group of all other ruminants, similar to previous analyses. Moschiola forms the sister-group to the other two tragulid genera Tragulus (from Asia) and Hyemoschus (from Africa), which is unexpected as usually the Asian species are thought to form a monophyletic group.

Complete mitochondrial genome sequence of Asiatic lion (Panthera leo persica).

The complete mitochondrial genome sequence 17,059 bp of Asiatic lion (Panthera leo persica) has been sequenced with the use of next generation sequencing technology using Ion Torrent PGM platform. The complete mitochondrial genome sequence of Asiatic lion consists of 13 protein-coding, 22 tRNA, and two rRNA genes, and 1 control region (CR). The mitochondrial genome is relatively similar to other felid mitochondrial genomes with respect to gene arrangement, composition, tRNA structures and skews of AT/GC bases to be typical of those reported for other mammals. The nucleotide composition of Asiatic lion mitogenome shows that there is more A-T% than G-C% on the positive strand as revealed by positive AT and CG skews. The overall base composition is 31.9% of A, 27.2% of C, 14.5% of G, and 26.2% of T. Most of the genes have ATA start codons, except ND1, COX2, ATP8, ATP6, ND4, and ND5 have ATG start codons.

Complete mitochondrial genome sequence of the Indian clouded leopard (Neofelis nebulosa).

The complete mitochondrial genome of sequence 16,859 bp of Indian clouded leopard (Neofelis nebulosa) has been sequenced using next generation sequencing technology Torrent PGM platform. The complete mitochondrial genome sequence of clouded leopard consists of 13 protein-coding, 22 tRNA, and two rRNA genes and a control region (CR). The mitochondrial genome is relatively similar to other felid mitochondrial genomes with respect to gene arrangement, composition, tRNA structures and skews of AT/GC bases to be typical of those reported for other mammals. The nucleotide composition of the genome shows that there is more A-T% than G-C% on the positive strand as revealed by positive AT and CG skews. The base composition of the mitochondrial genome of clouded leopard is as follows: A, 5362 bp (31.8%); C, 4560bp (27.0%); G, 2475 bp (14.6%); T, 4462 bp (26.4%). Most of the genes have ATG initiation codons, except ND1, ND2, ND3, ND4, ND6, and CYTB (ATA start codon).

Role of excipients and polymeric advancements in preparation of floating drug delivery systems.

Since decade or two, the development of floating drug delivery systems becomes a significant and novel tool as having low density than gastric content. There are various advanced polymers including chitosan, eudragit, etc., and excipients such as; pore forming agent, surfactants, etc. All of them are discussed briefly, and results are concluded from various reputed researches. We have discussed all natural and synthetic systems with their effect on the release and other parameters which are essential for the floating formulation development.

Detection of 40 bp insertion-deletion (INDEL) in mitochondrial control region among sambar (Rusa unicolor) populations in India.

BACKGROUND: The mitochondrial DNA (mtDNA) control region is extensively used in the phylogeography of species. We examined sequence variations in the mtDNA control region of sambar (Rusa unicolor) populations from the South, Central and North India. RESULTS: Most of the samples collected from the south India exhibited a 40 bp insertion in the mtDNA control region. This insertion was not observed in the North and Central Indian populations. CONCLUSION: This study provided a potential marker for molecular screening and identification of sambar populations in the form of a distinct 40 bp insertion. Some populations in South India did not exhibit this insertion. It indicates that there could be an ecological barrier that might be preventing the expansion of insertion-positive sambar population.

Pre-Historic and Recent Vicariance Events Shape Genetic Structure and Diversity in Endangered Lion-Tailed Macaque in the Western Ghats: Implications for Conservation.

Genetic isolation of populations is a potent force that helps shape the course of evolution. However, small populations in isolation, especially in fragmented landscapes, are known to lose genetic variability, suffer from inbreeding depression and become genetically differentiated among themselves. In this study, we assessed the genetic diversity of lion-tailed macaques (Macaca silenus) inhabiting the fragmented landscape of Anamalai hills and examined the genetic structure of the species across its distributional range in the Western Ghats. We sequenced around 900 bases of DNA covering two mitochondrial regions-hypervariable region-I and partial mitochondrial cytochrome b-from individuals sampled both from wild and captivity, constructed and dated phylogenetic trees. We found that the lion-tailed macaque troops in the isolated forest patches in Anamalai hills have depleted mitochondrial DNA diversity compared to troops in larger and continuous forests. Our results also revealed an ancient divergence in the lion-tailed macaque into two distinct populations across the Palghat gap, dating to 2.11 million years ago. In light of our findings, we make a few suggestions on the management of wild and captive populations.

A novel microsatellite (STR) marker for forensic identification of big cats in India.

India is the home to five of the eight majestic big cats of the world. The three major big cats namely, lion, tiger, and leopard are listed in the Schedule I of the Indian Wildlife Protection Act, 1972. Apart from the severe loss of the habitat, these are continuously facing the danger of extinction mainly due to poaching and hunting for their body parts, which are being greatly valued by apothecaries marketing traditional Chinese medicines. With the advent of polymerase chain reaction (PCR), DNA-based markers have emerged as major tools in the arena of wildlife forensics. Microsatellites (short tandem repeats, STRs) are markers of choice because of their polymorphic and co-dominant nature. These strictly follow the Mendelian inheritance and are highly reproducible. We have identified a new microsatellite (STR) locus Ple 46, which shows amplification in a species-specific manner (size of STR) in all the members of the family felidae studied here. This PCR-based, non-invasive method opens a new avenue to forensic identification of big cats.

Conservation of wild animals by assisted reproduction and molecular marker technology.

Wild animals are an integral component of the ecosystem. Their decimation due to abrupt natural calamities or due to gradual human intervention would be disastrous to the ecosystem and would alter the balance in nature between various biotic components. Such an imbalance could have an adverse effect on the ecosystem. Therefore, there is an urgent need to put an end to the ever increasing list of endangered species by undertaking both in situ and ex situ conservation using tools of modern biology, to ascertain the degree of genetic variation and reproductive competence in these animals. This review highlights the development and use of molecular markers such as microsatellites, minisatellites, mitochondrial control region, cytochrome b and MHC loci to assess the genetic variation in various Indian wild animals such as the lion, tiger, leopard and deer. The review also presents data on the semen profile of the big cats of India. Reproductive technologies such as cryopreservation of semen and artificial insemination in big cats are also highlighted.

Abscess mimicking pre-cervical and submandibular cystic hygroma in a newborn.

Pre-cervical inflammation is rarely seen in a newborn. Those swellings that are seen are usually congenital, such as vascular malformations, teratoma, dermoid cyst, thyroglossal cyst, cystic hygroma (abnormal lymphatic tissue), and very rarely, inflammation secondary to infection. Being able to differentiate between each condition is important because the course, treatment and prognosis for each condition are different. This case of a neck abscess in a newborn is reported due to its resemblance to cystic hygroma and highlights the similarities between the two conditions.

Urban poor children.

One-half of the world's population lives in cities and towns; this is expected to increase to 70% by 2050. One in three urban dwellers lives in slums. As the urban population grows, so does the number of urban poor. Out of a billion children living in urban areas, approximately 300 million are suffering from exclusion or are at risk of exclusion. Urban poor children are devoid of basic rights of survival, development and protection and are marginalised in challenging conditions in overcrowded settlements. Rapid urbanisation and the consequent increase in urban population is one of the biggest challenges that developing countries, including India are facing. Thirty per cent (that is, 367.5 million) of India's population of 1.23 billion live in urban areas. Moreover, this figure is increasing rapidly and is expected to reach 432 million (40%) by 2021. Rapid urbanisation has unfortunately outpaced development, and a large proportion (43 million) live in substandard conditions in slums. Now is the time to pay attention to the basic rights of the urban poor, especially the urban poor children, the most vulnerable group at the launching of 12(th) Five-Year Plan & National Urban Health Mission (NUHM) in India.

Integrated program achieves good survival but moderate recovery rates among children with severe acute malnutrition in India.

BACKGROUND: At any point in time, an average 8 million Indian children suffer from severe acute malnutrition (SAM). OBJECTIVE: This article assesses the effectiveness of an integrated model for the management of SAM (IM-SAM) in India comprising facility- and community-based care and using locally adapted protocols. DESIGN: Children (n = 2740) were randomly sampled from the 44,017 children aged 6-59 mo admitted to 199 Nutrition Rehabilitation Centers in the state of Madhya Pradesh (1 January to 31 December 2010). RESULTS: On admission, 2.2% of children had edema, 23.4% had medical complications, 56% were girls, 79% were in the age group 6-23 mo, and 64% belonged to scheduled tribe or scheduled caste families. Fifty-six children (2.0%) with severe congenital or pathological conditions were transferred to the district hospital. Of the 2684 program exits, 10 children (0.4%) died, 860 (32.0%) did not complete treatment (defaulted), and 1814 (67.6%) were discharged after a mean (±SD) stay of 75.8 ± 9.4 d. The mean weight gain among discharged children was 2.7 ± 1.9 g · kg body wt(-1) · d(-1); on discharge, 1179 (65%) of the children had recovered (weight gain ≥15% of initial weight). CONCLUSIONS: The survival rates in the IM-SAM program were very high. However, the moderate recovery rates documented seem to indicate that the protocols currently in use need to be improved. This trial was registered at clinicaltrials.gov as NCT01917734.

Species identification from dried snake venom.

Illegal trade in snake parts has increased enormously. In spite of strict protection under wildlife act, a large number of snakes are being killed ruthlessly in India for venom and skin. Here, an interesting case involving confiscation of crystallized dried snake venom and subsequent DNA-based species identification is reported. The analysis using the universal primers for cytochrome b region of the mitochondrial DNA revealed that the venom was extracted from an Indian cobra (Naja naja). On the basis of this report, the forwarding authority booked a case in the court of law against the accused for illegal hunting of an endangered venomous snake and smuggling of snake venom. This approach thus has immense potential for rapid identification of snake species facing endangerment because of illegal trade. This is also the first report of DNA isolation from dried snake venom for species identification.