Mullerian agenesis or Mayer-Rokitansky-Kuster-Hauser Syndrome (MRKH) Type-II is a congenital defect in the Mullerian duct that results in the absence of a uterus in women. The aetiology of this syndrome is unknown and has been considered a sporadic genetic disease. MRKH, together with anorectal anomaly, is an extremely rare condition and has only been reported in a few cases without any information on genetic analysis. This study investigated the mutational profile of a girl diagnosed with MRKH and anorectal anomalies with rectovaginal fistula. The whole exome sequencing (WES) trio-genetic analysis of a 5-year-old Malaysian girl diagnosed with MRKH (having anorectal anomaly with rectovaginal fistula) was performed together with her normal parents, using the Ion AmpliSeq Exome RDY kit (ThermoFisher Scientific, USA). Data were analysed using Torrent Suite v.5.0.4 and annotated using ANNOVAR. Single nucleotide polymorphisms (SNPs) with an allele frequency >0.01 were excluded, and the remaining variants were filtered based on de novo mutations, autosomal recessive, and autosomal recessive genetic traits. Related genes were analysed by biological pathway analysis (g:Profiler) and protein-protein interaction (HIPPIE v.2.3, STRING v.11.5, dan GeneMANIA). A total of 36 mutations were identified, and two of them, the LHX5 (p.P358Q), inherited from the father, and CFTR (p.R1158X), inherited from the mother. There were 28 de-novo mutations from 28 genes. All genes were involved in 27 biological processes that connected with 23 interactions, and are likely to cause MRKH syndrome in this patient.

Objective: This study examined the immune protection of immune-complex (Vaccine A) and attenuated live (Vaccine B) IBD vaccines in specific-pathogen-free (SPF) chickens against a novel Malaysian variant IBD virus (vaIBDV) challenge. Methods: One-day-old (n =75) SPF chickens were divided randomly into the following three groups of 25 chicks each: Control, Vaccine A, and Vaccine B groups. The vaIBDV strain, UPM1432/2019, was used for the challenge at 21 and 28days post-vaccination (dpv). Five birds from unchallenged and challenged groups were sacrificed seven days post-challenge, and blood, bursa, spleen, and cloacal swabs were collected. The IBD antibodies (Abs), lymphoid lesions, and viral load were determined. Results: The UPM1432/2019 virus induced bursal damage in vaccinated SPF chickens despite Ab titers. The mean Ab titers of the Vaccine A challenged group were significantly lower (p < 0.002) than in the unchallenged group at 28 dpv. The bursal indices of the vaccinated unchallenged groups did not differ significantly from those of the vaccinated challenged groups (p = 0.94). Microscopically, the bursae of the challenged groups showed significant atrophy. The bursal lesion score was higher (p < 0.05) in the control and Vaccine B challenged groups than the Vaccine A challenged group. The challenged group had a higher viral load than the vaccinated groups (p < 0.001). Conclusions and Relevance: Neither vaccine fully protected against a vaIBDV challenge, highlighting the limitations of current vaccines and the need for further research.

Aims: Hypoxia is believed to be one of the key components contributing to the clinical resistance of cancer therapies. Alternative strategies are under investigation to overcome this resistance and the oncolytic virus stands amongst the others. Newcastle disease virus (NDV) has been demonstrated to possess oncolytic activity against cancer cells. The present study investigated the effects of oncolytic NDV strain AF2240 and V4-UPM on osteosarcoma cells (Saos-2) under normoxic and hypoxic conditions. Methodology and results: Results showed that the NDV strain AF2240 and V4-UPM could infect and kill normoxic and hypoxic Saos-2 cells equally well by inducing hypoxia-independent apoptosis, and S-phase cell cycle arrest under the microscopy examination, cell viability assay, Annexin V apoptosis assay and cell cycle analysis experiments. However, the Velogenic NDV strain AF2240 excelled over the lentogenic NDV V4-UPM with increased oncolytic effects in Saos-2 cells. Conclusion, significance and impact of study In a nutshell, normoxia or hypoxia microenvironment has little effect on NDV-induced oncolysis of Saos-2 cancer cells which poses as a potential agent for the treatment of resistant cancer.

Background: The commercially available Newcastle disease (ND) vaccines were developed based on Newcastle disease virus (NDV) isolates genetically divergent from field strains that can only prevent clinical disease, not shedding of virulent heterologous virus, highlighting the need to develop genotype-matched vaccines Objectives: This study examined the efficacy of the NDV genotype-matched vaccine, mIBS025 strain formulated in standard vaccine stabilizer, and in carboxymethyl sago starch-acid hydrogel (CMSS-AH) following vaccination via an eye drop (ED) and drinking water (DW). Methods: A challenge virus was prepared from a recent NDV isolated from ND vaccinated flock. Groups of specific-pathogen-free chickens were vaccinated with mIBS025 vaccine strain prepared in a standard vaccine stabilizer and CMSS-AH via ED and DW and then challenged with the UPM/NDV/IBS362/2016 strain. Results: Chickens vaccinated with CMSS-AH mIBS025 ED (group 2) developed the earliest and highest Hemagglutination Inhibition (HI) NDV antibody titer (8log 2 ) followed by standard mIBS025 ED (group 3) (7log 2 ) both conferred complete protection and drastically reduced virus shedding. By contrast, chickens vaccinated with standard mIBS025 DW (group 5) and CMSS-AH mIBS025 DW (group 4) developed low HI NDV antibody titers of 4log 2and 3log 2 , respectively, which correspondingly conferred only 50% and 60% protection and continuously shed the virulent virus via the oropharyngeal and cloacal routes until the end of the study at 14 dpc. Conclusions The efficacy of mIBS025 vaccines prepared in a standard vaccine stabilizer or CMSS-AH was affected by the vaccination routes. The groups vaccinated via ED had better protective immunity than those vaccinated via DW.

Background: The role of Toll-like receptors (TLRs) in a feline infectious peritonitis virus (FIPV) infection is not completely understood. Objectives: This study examined the expression of TLR3, TLR7, TLR9, tumor necrosis factoralpha (TNF-α), interferon (IFN)-β, and interleukin (IL)-10 upon an FIPV infection in CrandellReese feline kidney (CRFK) cells and feline monocytes. Methods: CRFK cells and monocytes from feline coronavirus (FCoV)-seronegative cats and FCoV-seropositive cats were infected with type II FIPV-79-1146. At four, 12, and 24 hours postinfection (hpi), the expression of TLR3, TLR7, TLR9, TNF-α, IFN-β, and IL-10, and the viral load were measured using reverse transcription quantitative polymerase chain reaction. Viral protein production was confirmed using immunofluorescence. Results FIPV-infected CRFK showed the upregulation of TLR9, TNF-α, and IFN-βexpression between 4 and 24 hpi. Uninfected monocytes from FCoV-seropositive cats showed lower TLR3 and TLR9 expression but higher TLR7 expression compared to uninfected monocytes from FCoV-seronegative cats. FIPV-infected monocytes from FCoV-seropositive cats downregulated TLR7 and TNF-α expression between 4 and 24 hpi, and 4 and 12 hpi, respectively. IFN-β was upregulated early in FIPV-infected monocytes from FCoV-seropositive cats, with a significant difference observed at 12 hpi compared to FCoV-seronegative cats.The viral load in the CRFK and FIPV-infected monocytes in both cohorts of cats was similar over time.ConclusionTLR7 may be the key TLR involved in evading the innate response against inhibiting TNF-α production. Distinct TLR expression profiles between FCoVseronegative and FCoV-seropositive cats were observed. The associated TLR that plays a role in the induction of IFN-β needs to be explored further.

Aims: Symbiotic bee fungus associated with the stingless bee larval food has been revealed to play a major role in performing a mutual relationship with the host. The fungus is believed capable to produce crucial nutrients that are required for larval pupation. However, detailed information on the fungus identification isolated from the larval food of our native Indo-Malaya stingless bee, Heterotrigona itama (Hymenoptera: Apidae), is poorly understood. Hence, this study was conducted to determine the effect of fungus isolated from the stingless bee larval food on the survival of H. itama larvae and identify the isolated fungus using both morphological and molecular analyses. Methodology and results: Elisa plates designed with F-bottom type were used as artificial brood cells in a controlled condition. The eggs transferred to Elisa plates were kept in the incubator with 75%-100% humidity and 25 ± 2.5 °C of temperature. This study carried out in three different treatments: (1) microbes-free larval food, (2) fresh larval food and (3) microbes-free larval food with fungus supplement. Results showed that the survival of H. itama larvae depends on the presence of beneficial fungus with the highest survival rate (Treatment 2 = 85.71%). The absence of the beneficial fungus in the fluid food of H. itama showed the lowest survival rate (Treatment 1 = 37.14%). Microscopy analysis showed that the fungus had various forms, including unconjugated asci, round-shaped ascospore and pseudohyphae. The molecular characterisation of the isolated fungi was performed using 23S rRNA gene sequencing using universal primers ITS1 and ITS4. DNA barcoding of three isolated fungi confirmed all isolated fungi matched Panus lecomtei (Basidiomycetes; Poriales; Polyporaceae) with 99.70% of similarity. Conclusion, significance and impact of study This study provides information on the importance of whitish fungus in appearance that existed in the brood cell, which is proposed to be the crucial component of in-vitro stingless bee queen rearing protocol.
Bees--microbiology

Background: Inclusion body hepatitis (IBH) is an economically important viral disease primarily affecting broiler and breeder chickens. All 12 serotypes of fowl adenovirus (FAdV) can cause IBH. Objectives: To characterize FAdV isolates based on phylogenetic analysis, and to study the pathogenicity of FAdV-8b in specific-pathogen-free (SPF) chickens following virus inoculation via oral and intramuscular (IM) routes. Methods: Suspected organ samples were subjected to virus isolation and polymerase chain reaction (PCR) for FAdV detection. Hexon gene sequencing and phylogenetic analysis were performed on FAdV-positive samples for serotype identification. One FAdV-8b isolate, UPM/ FAdV/420/2017, was selected for fiber gene characterization and pathogenicity study and was inoculated in SPF chickens via oral and IM routes. Results: The hexon gene phylogenetic analysis revealed that all isolates belonged to FAdV-8b. The fiber gene-based phylogenetic analysis of isolate UPM/FAdV/420/2017 supported the grouping of that isolate into FAdV species E. Pathogenicity study revealed that, chickens infected with UPM/FAdV/420/2017 via the IM route had higher clinical score values, higher percent mortality, higher degree of the liver lesions, higher antibody response (p < 0.05), and higher virus shedding amounts (p < 0.05) than those infected via the oral route. The highest virus copy numbers were detected in liver and gizzard. Conclusions FAdV-8b is the dominant FAdV serotype in Malaysia, and pathogenicity study of the FAdV-8b isolate UPM/FAdV/420/2017 indicated its ability to induce IBH in young SPF chickens when infected via oral or IM routes.

Background: Inclusion body hepatitis (IBH) is an economically important viral disease primarily affecting broiler and breeder chickens. All 12 serotypes of fowl adenovirus (FAdV) can cause IBH. Objectives: To characterize FAdV isolates based on phylogenetic analysis, and to study the pathogenicity of FAdV-8b in specific-pathogen-free (SPF) chickens following virus inoculation via oral and intramuscular (IM) routes. Methods: Suspected organ samples were subjected to virus isolation and polymerase chain reaction (PCR) for FAdV detection. Hexon gene sequencing and phylogenetic analysis were performed on FAdV-positive samples for serotype identification. One FAdV-8b isolate, UPM/ FAdV/420/2017, was selected for fiber gene characterization and pathogenicity study and was inoculated in SPF chickens via oral and IM routes. Results: The hexon gene phylogenetic analysis revealed that all isolates belonged to FAdV-8b. The fiber gene-based phylogenetic analysis of isolate UPM/FAdV/420/2017 supported the grouping of that isolate into FAdV species E. Pathogenicity study revealed that, chickens infected with UPM/FAdV/420/2017 via the IM route had higher clinical score values, higher percent mortality, higher degree of the liver lesions, higher antibody response (p < 0.05), and higher virus shedding amounts (p < 0.05) than those infected via the oral route. The highest virus copy numbers were detected in liver and gizzard. Conclusions FAdV-8b is the dominant FAdV serotype in Malaysia, and pathogenicity study of the FAdV-8b isolate UPM/FAdV/420/2017 indicated its ability to induce IBH in young SPF chickens when infected via oral or IM routes.

Homocystineimia is an Inborn Errors of Metabolism (IEM) which can occur due to accumulation of homocysteine. Homocysteine is one of the sulfur-containing amino acid with thiol group that is formed by demethylation of methionine. Deficiency of enzymes involves in homocysteine metabolism can give rise to seven types of homocystinemia subject to total homocysteine level. Therefore reference ranges are needed to differentiate between normal and abnormal population as well as the type of homocystineimia depending on the enzymes defect in the pathway. Hence, homocysteine reference ranges in children for the Malaysian population were postulated. 3 mL of blood was collected from 86 normal individuals (52 boys and 34 girls) and then subsequently processed and analysed using High Performance Liquid Chromatogrphy – Ion Exchange Chromatography (HPLC-IEC). The calculated mean total homocysteine for the population was 8.1 ± 3.89 µM (95% confidence interval, l 7.3-8.9 µM). Reference range was 2.5 – 16.2 µM with lower and upper cut-off were 1.0 µM dan 21.0 µM, respectively. The newly developed reference range of total homocysteine for Malaysian children is able to reduce false negative cases in the laboratory.

Background: The predominant infectious bronchitis virus (IBV) strains detected in chickens in Malaysia are the Malaysian variant (MV) and QX-like, which are associated with respiratory distress, nephropathy, and high mortality. On the other hand, the antigenic relatedness and efficacy of IBV vaccines against these 2 field IBV strains are not well characterized. Objectives: This study aimed to determine the antigen relatedness and efficacy of different IB vaccine strains against a challenge with MV and QX-like strains. Methods: The antigen relatedness and the ability of different IB vaccine strains in conferring protection against MV and QX-like were assessed based on the clinical signs, macroscopic lesions, and ciliary activity. Results: The MV strain IBS037A/2014 showed minor antigenic subtype differences with the vaccine virus Mass H120 and 4/91 strains but showed major antigenic subtype differences with the K2 strain. The Malaysian QX-like strain IBS130/2015 showed major antigenic subtype differences with the MV strain IBS037A/2014 and the vaccine strains except for K2. Chickens vaccinated once with Mass (H120) or with non-Mass (4/91 and K2) developed antibody responses with the highest antibody titer detected in the groups vaccinated with H120 and 4/91. The mean ciliary activities of the vaccinated chickens were between 56 to 59% and 48 to 52% in chickens challenged with IBS037A/2014 and IBS130/2015, respectively. The vaccinated and challenged birds showed mild to severe lesions in the lungs and kidneys. Conclusions Despite the minor antigenic subtype differences, a single inoculation with Mass or non-Mass vaccines could not protect against the MV IBS037A/2014 and QX-like IBS130/2015.