Search

The present study was aimed at the identification, molecular characterization, and risk factor assessment of Theileria infection among sheep of Haryana province, north India. A total of 402 blood samples were collected from three different climatic zones of Haryana from March 2020 to September 2021. Light microscopy of blood smears revealed Theileria spp. infection in 47.26% (n = 190), while 60.94% (n = 245) of blood samples were positive using nested PCR. Extensive molecular characterization of Theileria infection using four pairs of species-specific primers indicated the dominance of T. ovis (29.1%) followed by T. lestoquardi (12.69%), T. luwenshuni (5.97%) and T. annulata (1.49%). Mixed infection was detected in 11.69% of cases. Bidirectional sequencing and phylogeny further confirmed the presence of these four Theileria spp. in the investigated area under study. Hematology indicated a significant (p < 0.01) reduction in various haematological indices of animals infected with T. luwenshuni and T. lestoquardi compared to the healthy control group. Risk factors like age, sex, and zone were significantly associated with Theileria infection in sheep. The present investigation depicts the first comprehensive molecular report of ovine Theileria spp., which warrants further study to develop suitable control strategies against these haemoparasitic infections. [ABSTRACT FROM AUTHOR]

In the rice agroecosystems of Southeast Asia, rice root knot nematode (Meloidogyne graminicola) significantly impairs yield, representing a major species within the 'graminis-group' known for its morphological similarities with other root knot nematodes (RKNs). This study delves into the variations in reproductive potential, morphology, morphometrics, and genetic diversity among thirty RKN populations in rice across three distinct agroecological zones in Jharkhand, India. Despite notable differences in reproductive potential among the populations, morphological and morphometric correlations to reproductive potential were inconclusive. However, male and juvenile morphometrics were crucial for identifying intraspecific variability. Genetic analysis utilizing five molecular markers (ITS, 18 S rRNA, D2-D3 of 28 S, COX-I, and COX-II) affirmed the populations as M. graminicola, with ITS marker revealing significant intraspecific variability. Phylogenetic analysis underscored the close relationship between M. oryzae and M. graminicola, distinct from other mitotic RKN species. Low genetic distance and nucleotide diversity, coupled with high haplotype diversity, negative Tajima's D, and Fu's Fs of haplotype network analysis, suggested that all M. graminicola populations are expanding. These findings highlight the urgent need for comprehensive management strategies against M. graminicola, providing valuable insights for growers, extension officials, and plant breeders to develop targeted management approaches and resistance breeding programs., Competing Interests: Declarations. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Purpose: Epidemiological studies on amoebic infections are complicated due to morphologically identical and clinically important Entamoeba species. Therefore, newer, simpler, and more economical diagnostic techniques are required for differentiating clinically important Entamoeba species., Methods: We developed a single-round multiplex PCR assay to identify E. histolytica, E. moshkovskii, E. dispar, E. bangladeshi, and E. coli. Primers were designed based on variations in 18 S rRNA sequences. Sensitivity and specificity were assessed using known positive and negative samples. Furthermore, we screened 472 diarrheal samples using this technique alongside the reference PCR method to evaluate its suitability for epidemiological studies and clinical diagnosis. DNA sequencing and phylogenetic analysis of the isolates were conducted. All statistical analyses of the data were performed using GraphPad Prism., Results: The designed primers successfully yielded species-specific PCR products of different sizes as expected. We did not observe any non-specific amplifications of the primer set. The diagnostic performance was also convincing. After screening clinical samples using the method, we observed that 2.33% (n = 11) tested positive for E. moshkovskii, 1.06% (n = 5) tested positive for E. histolytica, and 0.85% (n = 4) tested positive for E. bangladeshi in the studied area. DNA sequencing further confirmed the identified species. The constructed phylogenetic tree also demonstrated clear separation of the detected species lineages., Conclusion: The study suggests the multiplex PCR assay could be a reliable diagnostic tool for amoebic infections. This study is particularly significant as it marks the first reported occurrence of E. bangladeshi since its documentation in South Africa and its native Bangladesh., Competing Interests: Declarations. Ethical Approval: The ethical clearance of this study was reviewed and approved by the Institutional Human Ethics Committee of the ICMR-NICED. Informed consent statements were obtained from the participants. Competing Interests: The authors affirm that they do not have any competing interests related to the publication of the article., (© 2024. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Purpose: Theileriosis is an economically important tick-borne pathogen with a serious impact on livestock health and productivity. Despite the fact that bovine theileriosis has been widely investigated, there exists a paucity of information on these infections in small ruminants, especially in India. The present study was carried out to detect and differentiate different Theileria spp. in goats using nested PCR RFLP. Methods: Blood samples and ticks were collected from 405 goats in various agro-climatic zones of Haryana state, India. The blood samples were screened by microscopy, nested PCR–RFLP, and sequence analysis. The nested PCR–RFLP was performed with four restriction enzymes viz., Hpa II, Bsh 1285I, Hae II and Rsa I. Six nested PCR amplicons with different RFLP patterns were sequenced and submitted to NCBI (OM666861, MZ220430, OM666628, MZ220437, OM666637, OM721806). Results: Microscopy revealed 18.27% (n = 74) infection with Theileria spp., while 33.58% (n = 136) of blood samples were confirmed positive by nested PCR. Out of 136 positive samples, 43.38% (n = 59), 11.02% (n = 15) and 20.58% (n = 28), were positive for T. ovis, T. lestoquardi and T. luwenshuni (Theileria sp. China 1), respectively. Mixed infection was detected in 25% (n = 34) cases. Based upon Hpa II digestion pattern, 13 samples with T. lestoquardi and T. ovis, and 21 samples with T. ovis and T. luwenshuni were detected. Sequence study further confirmed their identity. The majority of ticks collected from goats were identified as Rhipicephalus spp., Hyalomma anatolicum and Hemaphysalis spp. Conclusion: This study represents the first confirmed molecular report of goats infected with T. ovis, T. lestoquardi, and T. luwenshuni from northern India. [ABSTRACT FROM AUTHOR]

Ovine theileriosis is an important tick-borne haemoprotozoan disease of sheep in tropical and subtropical regions, causing severe productivity and economic loss. There is a paucity of information related to molecular studies of ovine theileriosis from India. The present study identified different Theileria spp. in naturally infected sheep using nested PCR-restriction fragment length polymorphism (nPCR-RFLP). Blood samples and ticks were collected from 204 sheep in different agro-climatic zones of Haryana state, India, during the tick active season. Microscopic examination of thin blood smears revealed 33.3% (68/204) infections with Theileria spp., while 44.6% (91/204) of blood samples were positive by nPCR assay. Different Theileria spp. were identified based upon RFLP patterns using four restriction enzymes: Hpa II, Bsh 1285I, Hae II and Rsa I. Out of 91 positive samples, 50.5% (46/91), 23.08% (21/91), 11% (10/91) and 2.2% (2/91) were positive for T. ovis, T. lestoquardi, T. luwenshuni (Theileria sp. China 1/Theileria sp. China) and T. annulata, respectively. Mixed infection was detected in 13.2% (12/91) of cases. Based upon HpaII enzymatic digestion pattern, two samples with T. lestoquardi and T. annulata, nine samples with T. lestoquardi and T. ovis and one sample with T. ovis and T. annulata were detected. The presence of these Theileria spp. was further confirmed by sequence analysis. The majority of ticks collected from sheep were identified as Rhipicephalus spp. followed by Hyalomma anatolicum and Hemaphysalis spp. The present investigation depicts the first comprehensive molecular report of naturally infected sheep with T. ovis, T. lestoquardi, T. annulata and T. luwenshuni from northern India. [ABSTRACT FROM AUTHOR]

Summary: Equine babesiosis in India has always been a severe and life‐threatening disease attributed to infection with Theileria equi and Babesia caballi. These organisms are not host‐specific, as both T. equi and B. caballi are reported in dogs. Likewise, Babesia canis canis and Babesia canis rossi, the common causative agents in canines, have also been identified in horses. However, no molecular evidence of Babesia gibsoni has been reported in horses in India. A 5‐year‐old Thoroughbred gelding residing in Bengaluru showed signs of recurrent intermittent fever, inappetence, poor performance, anaemia and depression. A regular complete blood count and biochemical profile were done, however, the results were unresponsive to treatment interventions. The blood sample was submitted for screening equine haemoprotozoans using polymerase chain reaction. The current blood sample was screened for haemoprotozoans. The sample was positive for B. gibsoni and was confirmed by sequencing. Basic Local Alignment Search Tool (BLAST) search against GenBank revealed the highest similarity (95.17%, Accession No. MN385430.1) with B. gibsoni 18s rRNA partial sequence. [ABSTRACT FROM AUTHOR]

Background: The study was aimed at molecular detection and assessment of important biomarkers in the natural cases of canine babesiosis. Methods: Blood samples of 239 dogs were examined in PCR by targeting 18S rRNA gene. Hematobiochemical, oxidant-antioxidant and plasma cortisol parameters were estimated in the dogs on the day of presentation. Result: The 18S rRNA gene sequence showed 100% homology with Babesia canis vogeli and phylogram formed a tight cluster of B. canis vogeli originated from India/other countries. Higher prevalence rate (P<0.05) was noted in the PCR (7.95%) than the cytological technique (3.76%). Hemogram of infected dogs showed decrease (P<0.05) in the mean value of hemoglobin, RBC, WBC, HCT, whereas an increase in MCHC, lymphocytes, eosinophils, monocytes and thrombocytes. The ALT (49.29±1.53 U/L), AST (48.33±2.93 U/L), total protein (10.56±0.60 g/dL), creatinine (1.41±0.10 mg/dL) and urea (19.32±0.97 mg/dL) showed significant (P<0.005) increase, whereas decrease in the levels of serum glucose (82.76±2.78 mg/dL) in the infected dogs. Activity of MDA and SOD was significantly (P<0.01) increased (7.50±7.08 nmole/µL blood) and decreased (0.015±3.91 nmole/µL blood) in the diseased dogs, respectively. Plasma cortisol concentration was 11.10±7.84 nmol/L and 2.77±5.78 nmol/L (P<0.01) in the infected and uninfected dogs, respectively. [ABSTRACT FROM AUTHOR]

Background: Interspecific hybrids of rohu (Labeo rohita) and catla (Labeo catla) are common, especially in India due to constrained breeding. These hybrids must segregate from their wild parents as part of conservational strategies. This study intended to screen the hybrids from wild rohu and catla parents using both morphometric and molecular approaches., Methods & Results: The carp samples were collected from Jharkhand and West Bengal, India. The correlation and regression analysis of morphometric features are considered superficial but could be protracted statistically by clustering analysis and further consolidated by nucleotide variations of one mitochondrial and one nuclear gene to differentiate hybrids from their parents. Out of 21 morphometric features, 6 were used for clustering analysis that exhibited discrete separation among rohu, catla, and their hybrids when the data points were plotted in a low-dimensional 2-D plane using the first 2 principal components. Out of 40 selected single nucleotide polymorphism (SNP) positions of the COX1 gene, hybrid showed 100% similarity with catla. Concerning SNP similarity of the 18S rRNA nuclear gene, the hybrid showed 100% similarity with rohu but not with catla; exhibiting its probable parental inheritance., Conclusions: Along with morphometric analysis, the SNP comparison study together points towards strong evidence of interspecific hybridization between rohu and catla, as these hybrids share both morphological and molecular differences with either parent. However, this study will help screen the hybrids from their wild parents, as a strategy for conservational management., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

In the present study, tissue samples (tongue, esophagus and heart) were investigated from dromedary camels of India for identification and characterization of Sarcocystis spp. using histopathology, PCR and gene sequencing. Genomic DNA extracted from these tissue samples was used for PCR amplification of the cytochrome c oxidase subunit I gene (cox1) of Sarcocystis spp. and the partial sequence of small subunit ribosomal RNA (18S rRNA) gene of the S. cameli. The PCR products were purified, sequenced and analyzed using bioinformatics tools. Based on phylogenetic analysis of the cox1 gene, the sequences of the present study clustered with those of S. cameli, hosted by dromedary camels of Iraq and a close association was observed with S. masoni hosted by dogs and alpacas of China. Until now, there are no 18S rRNA sequences of S. cameli available in GenBank and this is the first study recording 18S rRNA sequences of S. cameli which were grouped with S. masoni from alpaca of China and guanaco and llama of Argentina in phylogenetic analysis. These findings could be useful for further studies on the characterization through molecular epidemiology, genetic diversity and host specificity of S. cameli., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Equine piroplasmosis caused by Theileria equi is a febrile, tick-borne disease of equids. However, there is limited literature about the genotyping of T. equi in India. Blood samples were collected from 202 horses and subjected to microscopy and PCR to detect T. equi. Initially, a universal screening primer pair targeting 18S ribosomal RNA genes common for Babesia caballi and T. equi was employed to amplify the DNA of both parasites. Thereafter additional primers were employed for species-specific detection resulting in amplification of approximately 435 bp specific for T. equi. T.equi was detected in 9.9% and 20.79% of horses screened by microscopy and PCR, respectively. The representative samples confirmed positive by PCR were sequenced, submitted to NCBI (OR651254, OR687254, OR685656, OR650830, OR650834), and used for genotype characterization and phylogenetic analysis. Employing Genetool and MEGA X software, the T. equi Indian isolates and across the globe were compared, and the results demonstrated 99.05-100% and 95.86-100% homologies, respectively. All the T. equi Indian isolates belonged to genotype A. Phylogeny based on the EMA-1 gene of five isolates (OR731831, OR731833, OR731829, OR731830, OR731832) were also characterized by sequencing and support the previous findings. Genotypes C and D, as well as genotypes B and E, exhibited lower levels of evolutionary divergence compared to other genotypes. The EMA-1 gene exhibited limited diversity and might not be the most suitable target for assessing variability within T. equi populations. The findings also reveal a significant association (p < 0.01) between T. equi infection and the presence of ticks., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A total of 57 tissue samples of domestic pigs (Sus scrofa) were collected from the meat outlets of five north Indian states and examined for sarcocystosis by histological and molecular methods. The genomic DNA extracted from five representative positive isolates was subjected to PCR amplification of the partial 18S rRNA gene followed by cloning and sequencing. Sequence analysis of the newly generated Indian isolates recorded 96.9–100.0% identity with published sequences of Sarcocystis suihominis. Two new haplotypes that have not been previously described manifested 99.5–100.0% nucleotide homology within themselves. In the phylogenetic analysis, Indian isolates of S. suihominis grouped together with S. suihominis originating from Italy, and they collectively formed a sister clade with Sarcocystis miescheriana within a clade containing various Sarcocystis spp. of ruminants having felids as final hosts. At the same time, this clade separated from a sister clade containing Sarcocystis spp. of bovid or cervid ruminants using canids as known or surmised definitive host. The current study established the phylogenetic relationship of Indian isolates of S. suihominis with various Sarcocystis spp. as well as with other taxa of Sarcocystidae family based on 18S rRNA gene for the first time. [ABSTRACT FROM AUTHOR]

Summary: A redescription of Distolabrellus veechi (Rhabditida: Mesorhabditidae) from agricultural soils in Uttarakhand and Uttar Pradesh, India, is provided. Specimens were extracted from soil using the Galleria soil baiting technique, with two strains of nematodes named JP1 and JP2 being obtained. Morphological and morphometric studies on the species agree well with previous records. Molecular analyses are provided using 18S, 28S and ITS rRNA sequences, the ITS sequences being obtained for the first time in this species. These analyses show that the material examined agrees well with previously studied populations. Phylogenetic analyses showed Distolabrellus to be the sister group of Crustorhabditis and Teratorhabditis , all having the male spicules fused for more than 50% of their length. Illustrations and phylogenetic trees based on 18S, 28S and ITS rRNA sequences are provided. [ABSTRACT FROM AUTHOR]

The flowerhorn cichlid is a popular ornamental fish in many Asian countries. The present study reports the occurrence of Paracapillaria philippinensis (Chitwood et al., 1968), a parasitic nematode in flowerhorn cichlid (Cichlasoma sp.) from south India. The infected fish demonstrated clinical symptoms viz. dark coloration, poor appetite, lethargy, erratic swimming, and white stringy faeces. Microscopic observation of the intestinal content and faeces revealed the presence of adult worms, larvae, and unembryonated eggs. PCR amplification of eukaryotic 18S rRNA, P. philippinensis-specific SSU rRNA gene, and the subsequent sequence analysis confirmed the species identity as P. philippinensis. The generated sequences were submitted in the GenBank, NCBI, under the accession numbers, MK895507.1, MK895446.1, MW144993.1, and OR685675.1. This is the first scientific report of P. philippinensis in fish from India, and it confirms that the flowerhorn cichlid can act as a definitive host for P. philippinensis. This report alerts fish handlers and enthusiasts to undertake suitable precautionary measures while handling live flowerhorn cichlids to prevent possible transmission of P. Philippinensis, which has the potential to infect humans causing intestinal capillariasis., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Microalgae can be an alternative to petroleum. In this paper, microalgal isolates were isolated from water samples of Tamil Nadu, India and were identified as Tetradesmus wisconsinensis and Botryococcus sp. by 18S rRNA neighbor-joining method. Nile red staining confirmed the presence of lipid in the cells. The total FAME (fatty acid methyl esters) content recorded for Botryococcus sp. was 39.11% followed by Tetradesmus wisconsinensis (35.23%). The major FAMEs observed in both the isolates were octadecadienoic acid methyl ester, octadecenoic acid methyl ester, and hexadecanoic acid methyl ester, which are responsible for biodiesel production. Biodiesel properties determined confirmed the potential of algal biodiesel as a substitute for diesel fuel. [ABSTRACT FROM AUTHOR]

Sarcocystis tenella is a common tissue coccidian parasite of sheep. It is reported worldwide with high prevalence rate ranging from 9 to 100%. However, there are very limited reports of this parasite from the Indian context and those reports are totally based on the morphology alone. When it comes to molecular characterization, such studies are absent from India. The present communication reports the first characterization study of S. tenella from India. 18S rRNA ribosomal gene and mitochondrial cytochrome c oxidase subunit I (cox1) genes were used for molecular characterization and phylogenetic analysis alongside standard histopathology of sarcocysts. Five Indian isolates were characterized for each gene, and respective sequences were submitted in the NCBI. Two haplotypes were noticed, both for the 18S rRNA and cox1 gene showing 99.8–100.0% and 99.7–100.0% nucleotide homologies within themselves, respectively. When compared with other sequences of S. tenella across the globe, the present isolates showed 93.3–99.9% nucleotide homology based on 18S rRNA gene and 95.2–99.8% nucleotide homology based on cox1 gene, respectively. In both the 18S and cox1 phylogenetic trees, respective sequences of S. tenella were placed with monophyletic cluster which was sister to a cluster comprising of sequences of S. gracilis and S. alces. [ABSTRACT FROM AUTHOR]

To date, the family Plagiopylidae comprises four genera: Plagiopyla, Lechriopyla, Pseudoplagiopyla and Paraplagiopyla. Despite its cosmopolitan distribution, only a few species currently comprise the genus Plagiopyla, and their data are often incomplete, whereas the other genera are monotypic. Indeed, these anaerobic ciliates are a somewhat challenging issue for protistologists: they have a cryptic lifestyle, they are not easily cultivable and they possess few and cryptic taxonomic features, which makes it extremely difficult to identify them at species level. In the present paper we provide a revision of the family Plagiopylidae based on morphological and molecular analyses, together with the description of two novel species of Plagiopyla, P. ramani sp. nov. and P. narasimhamurtii sp. nov. and a redescription of P. nasuta, all collected from the freshwater Kolleru Lake (Andhra Pradesh, India). We applied an integrative taxonomic approach to characterize these organisms, while introducing the description of novel morphological features not recorded in previous works. After critically revising the family Plagiopylidae, we propose the exclusion of the genus Paraplagiopyla from this family and that the genus Lechriopyla should be considered a junior synonym of Plagiopyla. We consequently propose L. mystax as Plagiopyla mystax comb. nov. [ABSTRACT FROM AUTHOR]

Despite its frequent presence in buffaloes, Sarcocystis buffalonis remains as one of the most under studied parasite. In the present study, isolates of S. buffalonis from,Mathura, Uttar Pradesh India were characterized for 18S rRNA (MF595842-MF595844), cox 1 (MG792800-MG792802), 28S rRNA (MH793418-MH793420) and ITS 1 (MH793421-MH793423) genes. Analysis revealed multiple haplotypes for each individual gene viz., 18S rRNA (three haplotypes), cox 1 (two haplotypes), 28S rRNA (two haplotypes) and ITS 1 (single haplotype). The studied Indian sequences showed variable homologies for individual gene loci viz., 18S rRNA (99.3–99.9%); cox 1 (99.8–100.0%); 28S rRNA (99.9–100.0%) and ITS 1 (100.0%) The phylogenetic association between S. buffalonis and closely related Sarcocystis spp. infecting buffaloes and cattle was delineated. All these gene loci placed S. buffalonis close to S. hirsuta. The study has generated A vital phylogenetic data about this erstwhile neglected parasite. [ABSTRACT FROM AUTHOR]

Canine babesiosis is a vector borne disease caused by intra-erythrocytic apicomplexan parasites Babesia canis (large form) and Babesia gibsoni (small form), throughout the globe. Apart from few sporadic reports on the occurrence of B. gibsoni infection in dogs, no attempt has been made to characterize Babesia spp. of dogs in India. Fifteen canine blood samples, positive for small form of Babesia , collected from northern to eastern parts of India, were used for amplification of 18S rRNA gene (∼1665 bp) of Babesia sp. and partial ITS1 region (∼254 bp) of B. gibsoni Asian genotype. Cloning and sequencing of the amplified products of each sample was performed separately. Based on sequences and phylogenetic analysis of 18S rRNA and ITS1 sequences, 13 were considered to be B. gibsoni . These thirteen isolates shared high sequence identity with each other and with B. gibsoni Asian genotype. The other two isolates could not be assigned to any particular species because of the difference(s) in 18S rRNA sequence with B. gibsoni and closer identity with B abesia occultans and B abesia orientalis. In the phylogenetic tree, all the isolates of B. gibsoni Asian genotype formed a separate major clade named as Babesia spp . sensu stricto clade with high bootstrap support. The two unnamed Babesia sp. (Malbazar and Ludhiana isolates) clustered close together with B. orientalis , Babesia sp. (Kashi 1 isolate) and B. occultans of bovines. It can be inferred from this study that 18S rRNA gene and ITS1 region are highly conserved among 13 B. gibsoni isolates from India. It is the maiden attempt of genetic characterization by sequencing of 18S rRNA gene and ITS1 region of B. gibsoni from India and is also the first record on the occurrence of an unknown Babesia sp. of dogs from south and south-east Asia. [ABSTRACT FROM AUTHOR]

Namalycastis jaya sp. n. (Polychaeta: Nereididae: Namanereidinae) is described from the southern coast of Kerala in southwest India. One important characteristic feature of the species is the lack of notochaetae in all parapodia, a characteristic that it shares with at least two other species, Namalycastis elobeyensis Glasby, 1999 and Namalycastis hawaiiensis Johnson, 1903. It differs from N. elobeyensis by virtue of its smaller antennae, unequal eye size, bilobed acicular neuropodial ligule and multi-incised pygidium rim. Moreover, it differs from N. hawaiiensis by having fewer teeth on the serrated blades of the sub-neuroacicular falciger in chaetiger 10, and by possessing finely serrated falcigers in posterior segments. Beyond morphological analyses, molecular phylogenetics was used for the first time for Namalycastis to support population monophyly and recognition of the new species. The analysis, using both mitochondrial and nuclear data, corroborated the morphological analysis in suggesting that our specimens represent an as yet undescribed species, Namalycastis jaya sp. n., which forms a monophyletic group among the sampled nereidid taxa. Finally, a taxonomic key for Namalycastis species recorded from the Indian region is provided. [ABSTRACT FROM AUTHOR]

Abstract: Dung samples were collected from dairy calves of south Indian states viz., Andhra Pradesh, Karnataka, Kerala, Tamil Nadu and union territory, Puducherry and are subjected to nested polymerase chain reaction (PCR) targeting 18S rRNA gene for detection of Cryptosporidium infection. Of the 459 dung samples screened 182 were found positive with a prevalence of 39.65%. Highest prevalence of Cryptosporidium was observed in Puducherry (86.67%) and lowest in Kerala (17.65%). Genotyping by PCR-restriction fragment length polymorphism (RFLP) and sequence analysis revealed the presence of all the four major Cryptosporidium species of cattle viz., Cryptosporidium andersoni, Cryptosporidium ryanae, Cryptosporidium parvum and Cryptosporidium bovis. C. andersoni was widely distributed in calves of Tamil Nadu, Karnataka and Puducherry whereas in Andhra Pradesh C. ryanae was the major species. Of the 64 samples subjected to PCR-RFLP, 39 (60.94%) could be classified as C. andersoni, 18 (28.13%) as C. ryanae, 4 (6.25%) as C. parvum and 3 (4.69%) were confirmed as C. bovis. The results were also confirmed by sequencing of 19 Cryptosporidium DNA samples. [Copyright &y& Elsevier]

The present study was conducted to elucidate the population genetic diversity and haplotype network of Theileria annulata based on all available nearly complete 18S rRNA gene sequences in the GenBank™. In total, 52 sequences of the nuclear 18S rRNA gene used to assess the relationship of T. annulata with their country of origin identified 34 haplotypes. Haplotype 4 was widespread, occurring in India, China, Turkey and Iran, while the remaining haplotypes were singleton and unique to one country. Haplotype 4 displayed numerous single haplotypes around it and the stellate shape of the network suggested a rapid population expansion. India exhibited the largest number of haplotypes (h = 25) followed by Turkey (h = 6), China (h = 4), and Iran and Italy (h = 1). No geographical clustering of haplotypes was recorded. Nucleotide diversity was the highest in the Turkish followed by the Indian and Chinese populations. Similarly, haplotype diversity was the highest in China followed by Turkey, and the lowest in India. The overall dataset exhibited a low nucleotide diversity (0.00253 ± 0.00035), but high haplotype diversity (0.917 ± 0.034). It suggested the presence of only minor differences (01–11 nucleotide) between haplotypes which was also evident from the haplotype network. A high level of genetic diversity was documented within the Indian, Chinese and Turkish populations of T. annulata , whereas little genetic differentiation was noticed among these populations with a very high level of gene flow. Analysis of molecular variance (AMOVA) of T. annulata sequences revealed higher genetic variation within countries (83.58%) as compared to the variation among countries (16.42%). Neutrality indices, viz., Tajima's D, Fu and Li's F, Fu's F s , and R 2, along with the unimodal mismatch distributions demonstrated a recent population expansion of T. annulata in India and the overall dataset. However, the non-significant values of Tajima's D, Fu and Li's F, and Fu's F s for the Chinese population along with a bimodal mismatch distribution signified a constant population size. For the Turkish population, the neutrality and mismatch distribution tests either indicated a constant or a slight increase in population size. The present study provides novel insights into the population genetics and haplotype network of T. annulata based on the 18S rRNA gene for the first time. [Display omitted] • First study on the population structure and haplotype analysis of T. annulata based on 18S rRNA gene. • Overall 34 haplotypes of T. annulata were identified based on the 18S rRNA gene. • Haplotype 4 was widespread, occurring in India, China, Turkey and Iran. • Remaining 33 haplotypes were singleton and unique to one country. • Haplotype diversity was the highest in China followed by Turkey and India. • Neutrality indices showed population expansion in India and the overall dataset. [ABSTRACT FROM AUTHOR]

Sarcocystosis is a significant meat borne coccidian disease with immense zoonotic potential. Sarcocystis fusiformis is the most prevalent Sarcocystis spp. affecting buffaloes across the globe. Most of the molecular characterization works on S. fusiformis are from Egypt and there is no record of such work from India. In the present study, 21 isolates of S. fusiformis from Northern India were characterized for 18S rRNA (MF595821-MF595841) and cox 1 (MF423105-MF423119 and MH899162-MH899167) genes. S. fusiformis was seen as a monophyletic sister group to S. cafferi on the phylogenetic tree comprising of different Sarcocystis spp. Both genes placed S. fusiformis close to those Sarcocystis spp. which have felids as definitive hosts in comparison to those with canids as definitive host. A total of 15 and 7 haplotypes were noticed for both the genes, respectively. The studied Indian isolates showed 99.1–100.0% and 99.2–100.0% nucleotide homologies within themselves for both the respective gene loci. Over all, cox 1 gene was found to be better in delineating the evolutionary phylogenetics in comparison to 18S rRNA gene. The findings are important from evolutionary point of view. • First report of phylogenetic analysis of S. fusiformis from India. • A total of 15 and 7 haplotypes were noticed for 18S and cox 1 genes, respectively. • Phylogenetic position of S. fusiformis with respect to other Sarcocystis spp. was calculated. • Genetic diversity was found to be 99.1-100.0% and 99.2-100.0% for both the loci. [ABSTRACT FROM AUTHOR]

Genetic characterization of Theileria species infecting bovines in India was attempted targeting the 18S ribosomal RNA region of the parasite. Blood samples of bovines (n = 452), suspected for haemoprotozoan infections, from 9 different states of the country were microscopically examined for Theileria species infection. Four Theileria spp. positive blood samples from each state were randomly utilized for PCR amplification of the 18S rRNA gene (approx. 1529 bp) followed by cloning and sequencing. The sequence data analysis of all the 36 isolates revealed that 33 isolates had high sequence similarity with published sequences of T. annulata, whereas 3 isolates (MF287917, MF287924 and MF287928) showed close similarity with published sequences of T. orientalis. Sequence homology within the isolates ranged between 95.8 and 100% and variation in the length of targeted region was also noticed in different isolates (1527–1538 nt). Phylogenetic tree created for T. annulata sequences revealed that a total of 24 Indian isolates formed a major clade and grouped together with isolates originating from countries like China, Spain, Turkey and USA. Remaining 09 isolates clustered in a separate group and were closely related to the TA5 isolate of T. annulata (a new genotype) originating from India and also with the isolates from East Asian countries like Japan and Malaysia. All the three T. orientalis isolates had minimal intraspecific variation (99–100% homology) amongst themselves. Further, in the phylogenetic analysis T. orientalis Indian isolates were found to cluster away from other 14 isolates of T. buffeli / sergenti / orientalis originating from different countries (Australia, China, Indonesia and Spain). However, these 3 isolates clustered together with the T. buffeli Indian isolate (EF126184). Present study confirmed the circulation of different genotypes of T. annulata in India, along with T. orientalis isolates. • 18S rRNA gene based phylogenetic analysis of Theileria species infecting bovines in India was targeted. • Study recorded circulation of T. annulata and T. orientalis in the area of study. • Genetic variation was recorded in T. annulata circulating in India. [ABSTRACT FROM AUTHOR]

Kleinstyla dorsicirrata (Foissner, 1982) Foissner et al., 2002. comb. nov. (basionym: Gastrostyla dorsicirrata) is a slightly flexible oxytrichid, measuring about 88-115 × 27-46 μm in life and possesses cortical granules. Kleinstyla dorsicirrata is the only oxytrichid known so far with incompletely fragmented dorsal kinety. Morphological and morphogenetic data recognise K. dorsicirrata as nonstylonychine oxytrichid. Molecular phylogeny of an Indian population was inferred using 18S rRNA gene sequences and was examined with respect to oxytrichids exhibiting variation in dorsal kinety fragmentation. Kleinstyla dorsicirrata clusters with Oxytricha lanceolata; this proximity is quite significant as both show deviation from typical oxytrichid fragmentation of dorsal kinety. Molecular phylogeny of Indian population confirms its nonstylonychine oxytrichid status., (© 2014 The Author(s) Journal of Eukaryotic Microbiology © 2014 International Society of Protistologists.)

Botryococcus braunii GUBIOTJTBB1 was isolated from a freshwater reservoir in Assam, India and its taxonomic identity was confirmed by 18S rRNA sequence analysis. Biofuel potential of the microalga strain was assessed from batch culture under laboratory conditions, based on its lipid content and energy value of the dried biomass. Total lipid of 57.14% and hexane extractable crude hydrocarbon of 52.6% were recorded maximum at 56 and 28 days respectively, which vary upon culture durations. The energy value (54.69 kJ/g) of the strain's sundried biomass was found higher than that of petroleum diesel fuel and nearly twice than other microalgae strains compared. The strain GUBIOTJTBB1 was found superior in terms of total lipid and hydrocarbon contents comparing to the previously reported Indian strains of B. braunii. With further improvements in growth, the strain could become an ideal feedstock for potential biofuel production in the prevailing climatic conditions of the region., (Copyright © 2013 Elsevier Ltd. All rights reserved.)