Partial Nucleotide Sequence Analysis Research Articles

Occurrence of canine parvovirus type 2c in diarrhoeic faeces of dogs in Kolkata, India.

Canine parvovirus-2(CPV-2) causes a highly contagious disease of dogs characterised by acute hemorrhagic gastroenteritis, lethargy, vomiting, fever and usually bloody or mucoid diarrhoea. In the present study, 41 faecal samples collected from dogs exhibiting the signs of fever, vomition, bloody or mucoid diarrhoea in Kolkata, India were screened by haemagglutination test and PCR for detection of capsid protein coding VP2 gene. The viral genotype was detected by multiplex PCR and analysis of partial VP2 gene nucleotide sequences of selected PCR products with bioinformatics tool. Thirteen (31.71%) samples were found positive with HA titre ≥ 32 whereas 28 (68.29%) samples were positive by PCR of VP2 gene indicating higher sensitivity of PCR. Highest occurrence of CPV-2 was observed in the age group of 1-6 months (80.65%) and non-descript breeds with no history of vaccination (85%). Three samples were antigenic type CPV-2a, rest were CPV-2b/CPV 2c. Six CPV sequences were found to be highly similar to published CPV 2c sequences in BLAST analysis revealing a maximum identity of 99-100% with other CPV-2c strains and clustered together with CPV-2c strains of India and other countries in phylogenetic analysis. The present study highlights the need for continuous monitoring of samples to detect gradual changes in circulating CPV-2 genotypes in India.

A molecular phylogeny of the Persian Gulf and the Gulf of Oman oyster species

Abstract The taxonomy of oysters along the northern coasts of the Persian Gulf and the Gulf of Oman is not well recognized. We present a phylogenetic analysis of oyster species in these regions. We combined morphological and molecular techniques to obtain the identity of oysters to the lowest taxonomic levels. Analysis of partial nucleotide sequences from mitochondrial cytochrome c oxidase subunit I (COI) was used for the phylogenetic evaluation. Based on our findings, Iranian samples nested within the genus Saccostrea and belonged to Saccostrea mordax and Saccostrea palmula clades. The shell morphology of the studied samples was variable, as in other rock oyster species. The examination of morphological features was in line with the molecular outcomes, but despite some similarities, Iranian S. palmula had well-developed and elongated chomata. The results also showed that S. mordax and S. palmula possessed significant relative abundance as dominant oysters in the Persian Gulf and the Gulf of Oman, respectively. Phylogenetic analysis revealed that Iranian samples of S. palmula formed a separate subclade from the Gulf of California and Panama samples, with large genetic distances (6–7%). Iranian specimens differed morphologically and genetically, suggesting that they could be a new species, although more research is needed.

Detection, characterization and virulence analysis of nucleopolyhedrovirus isolated from the cotton leafworm, Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae)

BackgroundIn the present study, detection, characterization and virulence analysis of a field collected nucleopolyhedrovirus isolated from the cotton leafworm, Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae) was carried out. The obtained isolate, named SpliNPV-YW, was collected from diseased S. littoralis larvae in El-Menoufia governorate, Egypt.ResultsTransmission electron microscopy showed the presence of typical occlusion bodies with average size of (1.06 × 1.19 µm). Upon digestion using two different endonucleases, PstI and ScaI, no clear difference was detected in the collected isolate (SpliNPV-YW) DNA genome pattern compared to the reference strain SpliNPV-AN1956. The evolutionary analysis of the polyhedrin gene's partial nucleotide sequence revealed that SpliNPV-YW isolate was closed and had a genetic origin with the NPV isolate SpliMNPV-A26-5 that belongs to group II NPVs with identity of 99.7%. The median lethal concentration (LC50) and the median lethal time (LT50) values were estimated for second and fourth larval instars of S. littoralis. The LC50 values were 2.8 × 104 OB/ml for second larval instar and 5.2 × 105 OB/ml for fourth larval instar after 10 days of treatment. Regarding the speed of killing of the viral isolate, the results showed that the LT50 value for the second instar larvae (LT50 = 5.5 days) was lower than that of the fourth instar larvae (LT50 = 6.2 days) at concentrations of 4.3 × 1010 (ob/ml) and 1.2 × 1011 (ob/ml) for second and fourth instar larvae, respectively.ConclusionsHost specificity and virulence characteristics make SpliNPV-YW isolate a good potential to be utilized as a candidate biopesticide for the control of S. littoralis population in Egypt.

Detection of a New Variant in Outbreak of Bovine Ephemeral Fever in Turkey, 2020

Background: In this study, partial nucleotide sequence analysis of the G gene was performed for the molecular characterization of the virus that caused the bovine ephemeral fever virus (BEFV) epidemic in Turkey in 2020. Phylogenetic analysis of these nucleotide sequences was performed with the virus nucleotide sequences of the epidemics seen in 2008 and 2012. These sequences were announced in GenBank. Phylogenetic analysis of these nucleotide sequences was performed with the virus nucleotide sequences of the epidemics seen in 2008 and 2012. Methods: The study was conducted in dairy cattle holdings located in Diyarbakır Sur, Çınar and Dicle regions in South-eastern Turkey in August-November 2020. The number of animals in the holdings consisted of 750 (n=750), 150 (n=150) and 200 (n=200) cattle, respectively. Result: Severe respiratory symptoms and high mortality in the affected animals were notable symptoms. As a result of the phylogenetic analysis, it was determined that the virus that caused the epidemic in Turkey in 2020 was formed by a new variant in the Turkey-2 group, which was similar to the Indian isolates, unlike the Turkey-1 group, which was close to the Middle East variants in 2008 and 2012 isolates.

Identification, serotyping and antimicrobial susceptibility of Riemerella anatipestifer isolated from ducks in Vietnam

Background:Septicemia caused by Riemerella anatipestifer (R. anatipestifer) is a serious problem in the duck industry worldwide, and it is currently one of the major concerns for duck farming in Vietnam..Aim:This study was conducted to identify the causative agent of septicemia in ducks in Vietnam. The antimicrobial susceptibility and serotypes of R. anatipestifer isolates were also determined to provide valuable information for disease treatment and vaccine development.Methods:Riemerella anatipestifer was isolated using blood agar and chocolate agar media. The commercial API 20NE microtest system and the partial nucleotide sequence analysis of the 16s rRNA were used to identify R. anatipestifer strains. Serotypes were determined by slide agglutination test using standard antisera against R. anatipestifer. The disk diffusion method was utilized to investigate the antimicrobial susceptibility of R. anatipestifer isolated strains.Results:A total of 408 samples were collected from ducks with typical symptoms of septicemia for R. anatipestifer isolation. Sixty-nine R. anatipestifer strains were identified. Serotyping results showed that 30 out of 69 bacterial strains were classified as serotypes 1, 6, 8, 10, and 20, with serotype 10 being the most prevalent. The antimicrobial susceptibility test revealed that 100% of the bacterial isolates were susceptible to Amoxicillin/clavulanic acid and Imipenem. On the contrary, the majority of R. anatipestifer strains were resistant to Nalidixic acid (89.9%), Streptomycin (75.4%), and Norfloxacin (72.5%).Conclusion:This is the first ever report in terms of identification, serotyping, and antimicrobial susceptibility tests of R. anatipestifer causing septicemia in ducks of Vietnam, providing useful scientific information for treatment as well as vaccine development to control the disease.

DETECTION OF HERPESVIRUSES IN PASSERINE BIRDS CAPTURED DURING AUTUMN MIGRATION IN SLOVENIA.

Herpesviruses (HVs) were detected by PCR in the cloacal swabs of 0.76% (4/525) clinically healthy free-living passerine birds from 32 different species captured in mist nets in Slovenia during the 2014 and 2017 autumn migrations. Herpesviruses were detected in the Eurasian Blackcap (Sylvia atricapilla), the Common Blackbird (Turdus merula), and the Eurasian Blue Tit (Cyanistes caeruleus). Phylogenetic analysis of partial DNA polymerase gene nucleotide sequences of the HV strains showed a distant relationship with other alphaherpesviruses of birds. In the phylogenetic tree, the HVs detected were clustered together with HV detected in Sulphur-crested Cockatoo and Neotropic Cormorants, as well as with known HVs such as gallid HV1, psittacid HV1 and HV2, and passerine HV1. Different sequences of HVs with relatively low identity were detected in our study, suggesting that different HVs were circulating in passerines sampled during the autumn migration in Slovenia.

Molecular detection of feline leukemia virus in clinically ill cats in Klang Valley, Malaysia.

Background and Aim:Feline leukemia virus (FeLV) is classified as Retroviridae gammaretrovirus. FeLV occurs worldwide, including Malaysia. Thus far, only one decade-old study on molecular characterization of Malaysian FeLV isolates exists, which resulted in a scarcity of updated information of current FeLV isolates circulating in Malaysia. This study was conducted to determine the status of FeLV in clinically ill cats and to study the molecular characterization and phylogenetic relatedness of the current isolates.Materials and Methods:Convenience sampling was performed in 20 cats from the Gasing Veterinary Hospital in Selangor. Plasma and saliva samples were collected from 15 clinically ill cats and 5 healthy cats subjected to one-step reverse transcription-polymerase chain reaction with primers targeting a highly conserved gene of U3-LTR-gag.Results:Two clinically ill cats’ plasma and saliva samples tested positive for FeLV RNA. Partial nucleotide sequencing and phylogenetic analysis revealed that the current isolates were 94-99% homologous to the previous Malaysian and Japanese FeLV isolates.Conclusion:Current FeLV isolates from this study displayed higher similarity with the previous Malaysian isolates, signifying that a similar FeLV strain circulated among the cat population in Selangor.

Circulation of classic and recombinant human astroviruses detected in South Africa: 2009 to 2014.

Astroviruses (AstVs) are associated with diarrhoeal and extra-intestinal infections in human, animal and avian species. A prevalence of 7% was reported in selected regions in SA while AstVs detected from clinical stool specimens were almost identical phylogenetically to strains identified in environmental and water samples. This study investigated the molecular diversity of astroviruses circulating between 2009 and 2014 in South Africa (SA). Astroviruses detected in stool specimens collected from hospitalised children were investigated retrospectively. Astroviruses were characterised using type-specific RT-PCR, partial nucleotide sequence analyses in ORF1 and ORF2 and whole genome sequencing. Different genotypes were compared with clinical features to investigate genotype-related associations. The Vesikari severity scale (VSS) was evaluated for scoring astrovirus diarrhoeal infections. Of 405 astroviruses detected, 49.9 % (202/405) were characterised into 32 genotypes comprising 66.3 % (134/202) putative-recombinants and 33.7 % (68/202) classic strains. No trends by year of collection, age or site were observed. Whole genome analysis in eight strains revealed that genotypes assigned by partial nucleotide sequence analyses to five astroviruses were incorrect. Bivariate analyses showed there were no significant associations between genotypes and clinical symptoms or severity of infection. A comparison of Vesikari parameters with astrovirus-positive proxy values demonstrated that Vesikari scores for duration of diarrhoea and admission temperatures would result in a milder infection rating in astrovirus-positive cases. Diverse genotypes co-circulated with putative-recombinants predominating. Astrovirus classification was complicated by the lack of a consistent characterisation system and reliable reference database. The VSS should be used cautiously to rate astrovirus diarrhoea. While surveillance in communities and out-patient clinics must be continued, screening for human astroviruses in alternate hosts is needed to determine the reservoir species.

Leishmania naiffi andlainsoni in French Guiana: Clinical features and phylogenetic variability.

In French Guiana, five species are associated with Cutaneous Leishmaniasis (CL). Though infections with Leishmania guyanensis, L. (V.) braziliensis and L. (L.) amazonensis have been extensively described, there are few available clinical and genetic data on L. (V.) lainsoni and L. (V.) naiffi. We determined the clinical and epidemiological features of all cases of CL due to L. (V.) naiffi and L. (V.) lainsoni diagnosed in French Guiana between 2003 and 2019. Phylogenetic analysis was performed by sequencing a portion of HSP70 and cyt b genes. Five cases of L. naiffi and 25 cases of L. lainsoni were reported. Patients infected by L. (V.) lainsoni were usually infected on gold camps, mostly along the Maroni river (60%), while L. naiffi was observed in French patients infected on the coast (100%). A high number of pediatric cases (n = 5; 20%) was observed for L. (V.) lainsoni. A mild clinical course was observed for all cases of L. (V.) naiffi. HSP70 and cyt b partial nucleotide sequence analysis revealed different geographical clusters within L. (V.) naiffi and L. (V.) lainsoni but no association were found between phylogenetic and clinical features. Our data suggest distinct socio-epidemiological features for these two Leishmania species. Patients seem to get infected with L. (V.) naiffi during leisure activities in anthropized coastal areas, while L. (V.) lainsoni shares common features with L. (V.) guyanensis and braziliensis and seems to be acquired during professional activities in primary forest regions. Phylogenetic analysis has provided information on the intraspecific genetic variability of L. (V.) naiffi and L. (V.) lainsoni and how these genotypes are distributed at the geographic level.

Genome constellations of rotavirus a isolated from avian species in Brazil, 2008-2015.

Rotaviruses are members of the family Reoviridae and are a common cause of acute diarrhea in many mammalian and avian species. They are non-enveloped icosahedral particles and their genome comprises 11 segments of double-stranded RNA, which encodes six structural proteins (VP1-4, VP6-7) and six nonstructural proteins (NSP1-6). Genotypes are defined based upon the diversity found in these genes and viral characterization plays a central role on epidemiological studies and prevention. Here we investigate the distribution of Brazilian RVAs genotypes in 8 chicken samples collected between 2008 and 2015 from different regions by RT-PCR, partial (Sanger) nucleotide sequencing and phylogenetic analysis from all rotavirus genes. Although the identified genotypes were typical from avian host species, when analyzed together, they form novel genetic constellations: G19-P[31]-I11-R6-C6-M7-A16-N6-T8-E10-H8 and G19-P[31]-I4-R4-C4-M4-A16-N4-T4-E4-H4. This study highlights that avian rotaviruses are widespread among commercial farms in Brazil, and the co-circulation of at least two different genomic constellations indicates that may present a way bigger genetic variability, that can be increased by the possible transmission events from other birds, lack of specific preventive measures, as well as the different viral evolution mechanisms.

Detection and molecular characterization of chicken infectious anaemia virus in poultry flocks in Haryana

Chicken infectious anaemia virus (CIAV) is an economically important pathogen affecting poultry industry worldwide. Commercial poultry flocks (26) suspected to be affected with chicken infectious anaemia (CIA) were processed for Polymerase Chain Reaction (PCR) using VP2 gene primers and characterized by partial gene sequencing. The PCR revealed 26.9% (7/26) poultry flocks positive for CIAV. Eleven PCR products of CIAV DNA were sequenced. The partial nucleotide sequence analysis of VP2 genes revealed that 11 field strains had 99.2 to 100% similarity among themselves and with the Indian strains. The VP2 gene sequences of 11 field strains showed 97.5 to 100% similarity to the field strains sequences reported from all over the world. On the basis of partial nucleotide sequencing analysis of VP2 gene, our findings suggest that the viral strains circulating in Haryana have similarity to other Indian strains.

Distribution of a devastating fungal pathogen in mangrove forests of southern Iran

An epidemic of canker and dieback was found on Avicennia marina (Forssk.) Vierth and Rhizophora mucronata Lam. in the mangrove forests located in three different regions (the Qeshm Island, Sirik Port and Khamir Port) of Hormozgan province, on the southern coast of Iran, Persian Gulf. For A. marina, the highest mean of disease incidence was estimated in the Sirik Port (60.6%), followed by the Qeshm Island (50.4%) and Khamir Port (45.2%). Based on a dieback ordinal rating scale from 0 to 5, disease severity had the highest mean in the Sirik Port (3.1), followed by the Qeshm Island (2.7) and Khamir Port (2.5). The species R. mucronata was more affected by the disease than A. marina with the mean disease incidence of 62.5% and the mean disease severity of 3.2. A fungus was consistently recovered from the symptomatic tissues which was identified as Neoscytalidium dimidiatum based on morphological and molecular studies. Phylogenetic analysis of partial nucleotide sequence of the internal transcribed spacer (ITS) region (ITS1–5.8S-ITS2) validated the identity of the putative pathogenic fungus. Koch's postulates were fulfilled using healthy detached branches of both mangrove species inoculated with mycelium plugs of N. dimidiatum. This study is the first report worldwide concerning the occurrence of canker and dieback on A. marina and R. mucronata caused by N. dimidiatum.

First Report of Shoot Blight on Fragrant Pear Caused by Pantoea ananatis in China

Fragrant pears are important economic plants in Xinjiang region, a major agricultural production area in China. In March 2017, a strange shoot blight disease on fragrant pear was initially observed in Kolar, Xinjiang. The incidence of the disease was up to 50% in certain orchards, and the total incidence area was approximately 26,000 ha of commercial fields. The disease developed from blossoms to shoots and larger branches. More disease was observed on old and wounded trees. Symptoms included wilted shoot and leaves. When infections spread to larger tree limbs, the bark of branches became darkened and water-soaked. After bark shaving, dark xylem tissue was visible. Shoot samples were collected from diseased plants (Pyrus sinkiangensis Yu) with characteristic symptoms. The shoot samples were surface sterilized with 75% ethanol and 1% sodium hypochlorite, rinsed three times in sterilized water, cut into small pieces, and then soaked in 10 ml of sterile phosphate buffered saline (PBS) for 30 min. The suspension was streaked onto a nutrient agar plate and incubated at 30°C. After 48 h, dominating light yellow, round flat colonies with smooth surfaces were picked and restreaked for purification. The bacteria were identified by polymerase chain reaction amplified 16S rRNA gene and further by multilocus sequence analysis of partial nucleotide sequences of the genes rpoB and infB using rpoB CM7-F/CM31b-R (1,000 bp) and infB 01-F/02-R (1,030 bp) primer pairs, respectively (Brady et al. 2008). BLAST analysis showed that the 16S rRNA (NCBI accession no. MK128507), rpoB (MK135821), and infB (MK135822) gene sequences shared 100, 100, and 99% similarity to that of Pantoea ananatis strains BSP9 (KF360069.1), 15320 (MH015163.1), and BRT98 (MH015108.1), respectively. In April 2018, 3-year-old fragrant pear trees were inoculated with the P. ananatis in a separated garden. The branches of trees were injured with cuts of 2 to 3 cm in the bark. Autoclaved cotton balls were soaked with bacterial suspensions of 10⁸ CFU/ml and attached to the wounds with Parafilm. The bacterial suspensions were inoculated on three trees, and an additional two trees were treated with PBS instead as a negative control. Three shoots were inoculated per tree. The trees were then grown under natural conditions. After 12 weeks, symptoms resembling bacterial shoot blight symptoms had developed on all inoculated shoots. Control trees did not develop symptoms. Reisolations were performed from symptomatic tissues, and isolates were identified as P. ananatis using the techniques for molecular identification as mentioned above. P. ananatis was initially discovered on pineapple causing fruitlet rot in the Philippines in 1928 (Serrano 1928), and it is regarded as an emerging pathogen based on the increasing number of reports of diseases occurring on previously unrecorded hosts in different parts of the world. Coutinho et al. (2002) reported similar symptoms including shoot wilt and die-back after the infection of Eucalyptus trees with P. ananatis. In 2017, isolation and identification of brown rot bacteria P. ananatis was reported on imported succulent plants in China (Li et al. 2017). To our knowledge, this is the first report of P. ananatis causing shoot blight on fragrant pear in China. This study expanded the host range and geographic location of P. ananatis as a plant pathogen and suggests that commercial fragrant pear producers should take early control actions to prevent spread of and reduce losses caused by the disease.

Detection and Phylogenetic Analysis of Herpesviruses Detected in Wild Owls in Slovenia.

Herpesvirus (HV) was detected using PCR in the organs of eight of 55 wild owls (14.5%) from seven species that were found dead in various locations in Slovenia between 1995 and 2015. HV was detected in three species: the Eurasian eagle owl (Bubo bubo), Ural owl (Strix uralensis), and long-eared owl (Asio otus). Phylogenetic analysis of partial DNA polymerase gene nucleotide sequences showed that the detected HVs are similar to the avian and mammal alphaherpesviruses. Two sequences were very similar to known bird HV sequences. One sequence was identical to the columbid herpesvirus 1 (CoHV1) sequence, and the other was very similar to the gallid herpesvirus 2 (GaHV2) sequence. The phylogenetic tree revealed a lower similarity of the other six analyzed Slovenian sequences with the sequences of alphaherpesviruses of birds and mammals. This is the first study to report the detection of different HVs in owls.

Molecular identification of Aphidius transcaspicus (Hymenoptera: Braconidae: Aphidiinae), a closely associated species of Aphidius colemani, in Korea

Abstract Aphidius colemani (Braconidae: Aphidiinae) is an endoparasitoid and one of the most commercially used parasitoids for the control of various pest aphids. However, species identification of A. colemani is difficult because of the high degree of morphological similarity with closely associated species, such as A. transcaspicus and A. platensis. Recently, we identified a large-scale emergence of parasitoids from mummies of Rhopalosiphum maidis on barley banker plants in Yecheon County, Korea. The morphological characteristics of the forewing, petiole, and antenna indicated that our specimen is A. transcaspicus. The partial nucleotide sequence analysis of the mitochondrial cytochrome oxidase subunit I (COI) gene showed that it is 100% identical to Japanese specimens and 1.69% variable to Greek specimens. Analysis of the phylogenetic tree, haplotypes, and amino acid sequence of the COI gene, as well as the internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA region, indicated that A. transcaspicus is closer to A. platensis than A. colemani, and Korean and Japanese populations were mostly diversified groups at the intraspecific level. This study provides important information to elucidate the genetic variation in A. transcaspicus and the relationships between closely related species within the A. colemani group.

White-Tailed Sea Eagle (Haliaeetus albicilla) Die-Off Due to Infection with Highly Pathogenic Avian Influenza Virus, Subtype H5N8, in Germany.

In contrast to previous incursions of highly pathogenic avian influenza (HPAIV) H5 viruses, H5N8 clade 2.3.4.4b viruses caused numerous cases of lethal infections in white-tailed sea eagles (Haliaeetus albicilla) affecting mainly young eagles (younger than five years of age) in Germany during winter 2016/2017. Until April 2017, 17 HPAIV H5N8-positive white-tailed sea eagles had been detected (three found alive and 14 carcasses) by real-time RT-PCR and partial nucleotide sequence analyses. Severe neurological clinical signs were noticed which were corroborated by immunohistopathology revealing mild to moderate, oligo- to multifocal necrotizing virus-induced polioencephalitis. Lethal lead (Pb) concentrations, a main factor of mortality in sea eagles in previous years, could be ruled out by atomic absorption spectrometry. HPAIV H5 clade 2.3.4.4b reportedly is the first highly pathogenic influenza virus known to induce fatal disease in European white-tailed see eagles. This virus strain may become a new health threat to a highly protected species across its distribution range in Eurasia. Positive cloacal swabs suggest that eagles can spread the virus with their faeces.

Actinomycetes mediated targeting of drug resistant MRSA pathogens

The present study was to check the anti-Methicillin Resistant Staphylococcus aureus (MRSA) activity of actinomycetes isolated from terrestrial soil samples. The cell free supernatant and the ethyl acetate extract prepared from the potential isolate were studied for anti-MRSA activity. The culturing conditions of the potential isolate were optimized and characterized by polyphasic approach. The molecular taxonomy and phylogenetic studies of the potential isolate showed that it belonged to the genus Streptomyces. The partial 16S rDNA nucleotide sequencing analysis of the isolate yielded 1367 nucleotides and it was submitted to the GenBank under the accession number MF974561. Blast search of the nucleotide sequence of the isolate in the GenBank database showed 100% sequence similarity with Streptomyces gancidicus. The ethyl acetate (EA) extract showed 20 and 21 mm zone of inhibition against two MRSA strains (ATCC 43300) and (ATCC700699), respectively. Partial purification and GC-MS analysis of EA extract showed the presence of 1,1-Dichloropentane (DCP) (76%) as a major compound.

Sequence typing of human adenoviruses isolated from Polish patients subjected to allogeneic hematopoietic stem cell transplantation – a single center experience

ABSTRACT Purpose: Human adenoviruses (HAdV) from species A, B and C are commonly recognized as pathogens causing severe morbidity and mortality in hematopoietic stem cell transplant (HSCT) recipients. The purpose of the present study was to determine HAdV types responsible for viremia in HSCT recipients at a large tertiary hospital in Poland. Methods: Analysis of partial nucleotide sequences of HAdV hexon gene was used to type 40 clinical isolates of HAdV obtained from 40 HSCT recipients. Results: We identified six different HAdV serotypes belonging to species B, C and E. We demonstrated high variability in sequences of detected HAdV types, and patients infected with the same HAdV types were not hospitalized at the same time, which suggests the low possibility of cross-infection. In almost all patients, anti-HAdV antibodies in IgG class were detected, which indicates a history of HAdV infection in the past. Clinical symptoms accompanying HAdV viremia were in 89%, and in 61.5% of individuals, HAdV was a sole pathogen detected. There were no cases with high-level HAdV viremia and severe systemic or organ infections. Graft-versus-host disease (GvHD) was present in patients infected with species B and C, but grade II of GvHD was observed only in patients infected with HAdV-B. Conclusions: The predominance of HAdV-C and common presence of anti-HAdV antibodies in IgG class may strongly suggest that most infections in the present study were reactivations of HAdV persisting into the patient’s mucosa-associated lymphoid tissues. Variability of HAdV sequences suggests that cross-infections between patients were very rare. ABBREVIATIONS: GvHD: graft-versus-host disease; HAdV: human adenoviruses; HSCT: hematopoietic stem cell transplantation

Isolation and screening of Lactobacillus species from dogs for probiotic action

The present study aimed at the detection of suitable Lactobacillus species from dogs for usage as probiotic. A total of 67 rectal swabs from healthy pups were analyzed and 49 (73.1%) Lactobacillus isolates were identified based on morphological, biochemical characteristics and confirmed by genus specific PCR targeting 16S rRNA gene. A total of 20 isolates that showed strong aggregation, high cell surface hydrophobicity, acid and bile tolerance were screened for in-vitro antibacterial activity by agar well diffusion assay, where prominent inhibitory zones were observed against majority of the test pathogens. Reduction in antibacterial activity was noticed after neutralization, proteinase K and heat treatment of supernatants. Partial 16S rRNA nucleotide sequence analysis of seven Lactobacillus isolates that showed prominent in-vitro antibacterial activity revealed maximum sequence homology with Lactobacillus fermentum (for six isolates) and Lactobacillus agilis (for one isolate). The in-vitro antibacterial activity results highlighted the need for in-vivo studies in India to establish probiotic potential of canine faecal Lactobacillus species in the near future.

Noroviruses and sapoviruses associated with acute gastroenteritis in pediatric patients in Thailand: increased detection of recombinant norovirus GII.P16/GII.13 strains.

Enteric caliciviruses, including noroviruses (NoVs) and sapoviruses (SaVs), are recognized as important etiologic agents of acute gastroenteritis (AGE) with considerable genetic diversity. In order to gain an overview of the molecular epidemiology of human NoVs and SaVs in children hospitalized with AGE in Chiang Mai, Thailand, a total of 889 fecal specimens were collected from 2012 to 2014 and screened for NoVs and SaVs. Out of 889 fecal specimens, 154 (17.3%) and 6 (0.7%) were positive for NoV GII isolates and SaV, respectively. Among the NoV GII, 10 different genotypes were identified with genotype GII.4 being predominant (103 strains), followed by GII.3 (17 strains), GII.13 (13 strains), GII.1 (7 strains), GII.6 (7 strains), GII.7 (2 strains), GII.17 (2 strains), and one each of GII.2, GII.15, and GII.21 genotypes. It was observed that four variants of NoV GII.4 (Den Haag 2006b, Apeldoorn 2007, New Orleans 2009, Sydney 2012) were detected from 2012 to 2014. Analysis of partial nucleotide sequences of RdRp and VP1 of the emerging NoV GII.13 strains (9 of 13 strains) revealed that they all were GII.P16/GII.13 recombinants. In addition, four different genotypes of SaV, GI.1 (2 strains), GII.1 (1 strain), GII.4 (2 strains), and GIV.1 (1 strain) were detected. The data revealed heterogeneity and a highly dynamic distribution of NoV and SaV genotypes circulating in children admitted to hospitals with AGE in Chiang Mai, Thailand, during the period of 2012 to 2014.