Investigation of koala retrovirus in captive koalas with pneumonia and comparative analysis of subtype distribution.

This study focused on the involvement of koala retrovirus (KoRV) in pneumonia in koalas. Three deceased pneumonic koalas from a Japanese zoo were examined in this study. Hematological and histopathological findings were assessed, and KoRV proviral DNA loads in the blood and tissues were compared with those of eight other KoRV-infected koalas from different zoos. Demographic data and routine blood profiles were collected, and blood and tissue samples were analyzed to rule out concurrent infections in pneumonic koalas. KoRV subtyping and measurement of the KoRV proviral DNA load were performed by polymerase chain reaction (PCR) using specific primers targeting the pol and env genes. The results showed that the koalas had histopathologically suppurative and fibrinous pneumonia. Chlamydiosis was not detected in any of the animals. PCR analysis revealed KoRV-A, -B, and -C infections in all koalas, except for animals K10-11, which lacked KoRV-B. Significant variations in the proviral DNA loads of these KoRV subtypes were observed in all tissues and disease groups. Most tissues showed reduced KoRV loads in koalas with pneumonia, except in the spleen, which had significantly higher loads of total KoRV (2.54 × 107/µg DNA) and KoRV-A (4.74 × 107/µg DNA), suggesting potential immunosuppression. This study revealed the intricate dynamics of KoRV in various tissues, indicating its potential role in koala pneumonia via immunosuppression and opportunistic infections. Analysis of the levels of KoRV proviral DNA in different tissues will shed light on viral replication and the resulting pathogenesis in future studies.

Role of Clove and Tulsi on broiler health and meat production.

The role of Clove (Cv) and Tulsi (Ts) supplementation on broiler growth performance and gut health as an alternative to antibiotic growth promoters has already been established. Therefore, the objectives of this study were to investigate the role of Cv and Ts on the serum biochemical profile and meat quality traits in broilers. A total of sixty (60) one-day-old commercial broiler chicks were randomly allotted into four homogenous groups (15 birds per group). They were then fed Cv powder and Ts extract from day (d) 8 to d 28 with drinking water i.e. zero level of Cv or Ts (T0), 0.5% Cv + 2% Ts (T1), 1.0% Cv + 3% Ts (T2), 1.5% Cv + 4% Ts (T3). Blood and meat samples were collected on d 14, 21, and 28 to analyze the serum-biochemical profile and meat quality. Supplementation of Cv and Ts improved serum-biochemical profile by reducing total cholesterol and low-density lipoprotein levels in broilers. However, they did not affect the serum protein levels while the 0.5-1% Cv and 2-3% Ts supplemented groups had higher glucose levels on d 21. Production of breast and thigh meat increased with increased size, and density of myofibers while fed 0.5-1% Cv and 2-3% Ts. On the contrary, 1.5% Cv powder and 4% Ts extract supplementation improved thigh meat color, breast and thigh meat pH as well as the water-holding capacity. The current study findings suggest that Cv and Ts can be used in combination to improve broiler health, production, and meat quality.

Coagulation-adsorption-oxidation for removing dyes from tannery wastewater.

Dye removal from tannery wastewater is now a great concern given the ramifications for the environment in which the effluent ends up. Recently, the application of tannery solid waste as a byproduct to remove pollutants from tannery wastewater has garnered increasing attention. This study aims to extract biochar from tannery liming sludge for the removal of dye from wastewater. The activated (600 ºC) biochar was characterized by SEM (Scanning Electron Microscope), EDS (Energy Dispersive Spectroscopy), FTIR (Fourier Transform Infrared Spectroscopy), and surface area analysis utilizing the BET (Brunauer-Emmett-Teller) method and pHpzc (point of zero charges) analysis. The surface area and pHpzc of the biochar were determined as 9.29 m2/g and 8.7, respectively. The batch-wise coagulation-adsorption-oxidation was investigated for its efficacy in dye removal. The optimized conditions were as follows: the efficiency of dye, BOD (Biochemical Oxygen Demand) and COD (Chemical Oxygen Demand) were attained at 94.9%, 95.7% and 93.5%, respectively. SEM, EDS, and FTIR analyses before and after adsorption revealed that the derived biochar could adsorb dye from tannery wastewater. The adsorption behavior of the biochar fitted well to the Freundlich isotherm (R2 = 0.9987) and Pseudo 2nd order (R2 = 0.9996) kinetic model. This investigation provides a new dimension for state-of-the-art utilization of tannery solid waste as a feasible strategy to remove dye from tannery wastewater.

Effect of supplementary feeding on the production traits, carcass and meat quality of Jamuna basin lambs.

This study aimed to identify the optimum level of supplementary feeds on the carcass traits and meat quality of Jamuna basin lambs. Forty selected lambs were divided into four treatments such as T0 (no concentrate supplementation), T1 (1% concentrate feed), T2 (1.5% concentrate feed) and T3 (2% concentrate feed) having ten lambs per treatment. The data were analyzed through Completely Randomized Design (CRD) with SAS software. Hot carcass, dressing percentage, head, leg, neck, loin, heart, and spleen weight were showed significantly (p < 0.05) higher values with increasing concentrate feed. The crude protein (CP), ether extract (EE) and ash values were significantly increased (p < 0.001) except T2 treatment. The ultimate pH was significantly increased except T2 and cooked pH was significantly decreased (p < 0.001) except T3 treatment. Drip loss and cooking loss (CL) % had significantly reduced (p < 0.001) except T3 treatment. The water holding capacity (WHC) % was significantly increased (p < 0.001) except T3 treatment. The score of color, juiciness and tenderness were significantly different (p < 0.001). Flavor and overall acceptability score were significantly increased (p < 0.05) in different treatments. The color values L* and b* had significantly changed (p < 0.001) and a* value was found significantly higher (p < 0.05) in all treatments. Hence, 12 months of aged lambs with 1.5% concentrate feed showed better performances on carcass, nutritional, physicochemical, sensory and instrumental color values to increase the carcass and the meat quality of lambs.

Composting of limed fleshings generated in a tannery: sustainable waste management.

In tanneries, limed fleshing is an unavoidable waste generated in beamhouse operation. Proper management of limed fleshing with protein, fat, lime, and sulfide will help to protect the natural environment and at least reduce the pollution that ends up in it. In this study, excluding any pretreatment, limed fleshing is used for compost production. Chopped and mixed limed fleshing with chicken manure, cow dung, and sawdust was heaped onto a horizontal bamboo frame. Three composting heaps were fabricated weighing 720, 700, and 760 kg. The turning of composting materials in the heaps causes temperature changes in the thermophilic range. The thermophilic temperatures in these heaps were 69.07 °C (heap 1), 69.9 °C (heap 2), and 69.19 °C (heap 3) which ensured the death of the pathogenic organism. The quality of compost was assessed based on the nutrients-nitrogen (N), phosphorous (P), potassium (K), and sulfur (S) content. NPKS in the compost fulfils the requirements of the investigated materials as compost. The largest amounts of metals- zinc (Zn), copper (Cu), chromium (Cr), lead (Pb), and nickel (Ni) of the compost detected in the heaps were, respectively, 200.3, 37.4, 20.3, 12.0, and 3.9 mg/kg. Cadmium (Cd) in the compost was below the detection limit. Scanning electron microscope (SEM) photographs show the decomposing of composting materials. This study indicates that limed fleshing can be converted into nutrient-enriched compost without any pretreatment. Using an easy, simple, and adaptable technique could reduce the volume of solid waste generated in the tannery to reduce environmental pollution.

L-Carnitine Supports the In Vitro Growth of Buffalo Oocytes.

This study aimed to determine the effect of L-carnitine on the growth and subsequent nuclear maturation of buffalo small growing oocytes (92−108 µm in diameter) in vitro. Oocyte-granulosa cell complexes (OGCs) were dissected from early antral follicles of slaughtered buffaloes and cultured in in vitro growth (IVG) medium with the supplementation of different concentrations (0, 1.25, 1.875 or 2.5 mM) of L-carnitine for 6 days. The results revealed that L-carnitine increased the diameter of buffalo oocytes in vitro. The degeneration rate was significantly (p < 0.05) lower in 2.5 mM of L-carnitine-treated oocytes (10%) than others (55%, 45% and 32.5% in 0, 1.25 and 1.875 mM of L-carnitine-supplemented groups, respectively). The OGCs showed antrum-like structures significantly (p < 0.05) higher in the 2.5 mM of L-carnitine group (74.0%) than the 0- and 1.25-mM groups (34.6% and 38.1%, respectively). Furthermore, in vitro grown oocytes were placed in in vitro maturation (IVM) medium for 24 h to examine meiotic competence of in vitro grown oocytes with L-carnitine. The L-carnitine (1.875 and 2.5 mM) treated oocytes showed a higher rate of nuclear maturation up to the metaphase II (MII) stage and a lower rate of degeneration. In conclusion, L-carnitine enhances the growth, prevents degeneration, promotes the formation of antrum-like structures and supports nuclear maturation of buffalo oocytes in vitro.

Molecular Insights into Innate Immune Response in Captive Koala Peripheral Blood Mononuclear Cells Co-Infected with Multiple Koala Retrovirus Subtypes.

Koala retrovirus (KoRV) exists in both endogenous and exogenous forms and has appeared as a major threat to koala health and conservation. Currently, there are twelve identified KoRV subtypes: an endogenous subtype (KoRV-A) and eleven exogenous subtypes (KoRV-B to -I, KoRV-K, -L, and -M). However, information about subtype-related immune responses in koalas against multiple KoRV infections is limited. In this study, we investigated KoRV-subtype (A, B, C, D, and F)-related immunophenotypic changes, including CD4, CD8b, IFN-γ, IL-6, and IL-10 mRNA expression, in peripheral blood mononuclear cells (PBMCs) obtained from captive koalas (n = 37) infected with multiple KoRV subtypes (KoRV-A to F) reared in seven Japanese zoos. Based on KoRV subtype infection profiles, no significant difference in CD4 and CD8b mRNA expression was observed in the study populations. Based on the different KoRV subtype infections, we found that the IFN-γ mRNA expression in koala PMBCs differs insignificantly (p = 0.0534). In addition, IL-6 and IL-10 mRNA expression also did not vary significantly in koala PBMCs based on KoRV subtype differences. We also investigated the Toll-like receptors (TLRs) response, including TLR2-10, and TLR13 mRNA in koala PBMCs infected with multiple KoRV subtypes. Significant differential expression of TLR5, 7, 9, 10, and 13 mRNA was observed in the PBMCs from koalas infected with different KoRV subtypes. Therefore, based on the findings of this study, it is assumed that co-infection of multiple KoRV subtypes might modify the host innate immune response, including IFN-γ and TLRs responses. However, to have a more clear understanding regarding the effect of multiple KoRV subtypes on host cytokines and TLR response and pathogenesis, further large-scale studies including the koalas negative for KoRV and koalas infected with other KoRV subtypes (KoRV-A to -I, KoRV-K, -L and -M) are required.

In vivo Delivery Tools for Clustered Regularly Interspaced Short Palindromic Repeat/Associated Protein 9-Mediated Inhibition of Hepatitis B Virus Infection: An Update.

Chronic hepatitis B virus (HBV) infection remains a major global health problem despite the availability of an effective prophylactic HBV vaccine. Current antiviral therapies are unable to fully cure chronic hepatitis B (CHB) because of the persistent nature of covalently closed circular DNA (cccDNA), a replicative template for HBV, which necessitates the development of alternative therapeutic approaches. The CRISPR/Cas system, a newly emerging genome editing tool, holds great promise for genome editing and gene therapy. Several in vitro and/or in vivo studies have demonstrated the effectiveness of HBV-specific clustered regularly interspaced short palindromic repeat (CRISPR)/associated protein 9 (CRISPR/Cas9) systems in cleaving HBV DNA and cccDNA. Although recent advances in CRISPR/Cas technology enhance its prospects for clinical application against HBV infection, in vivo delivery of the CRISPR/Cas9 system at targets sites remains a major challenge that needs to be resolved before its clinical application in gene therapy for CHB. In the present review, we discuss CRISPR/Cas9 delivery tools for targeting HBV infection, with a focus on the development of adeno-associated virus vectors and lipid nanoparticle (LNP)-based CRISPR/Cas ribonucleoprotein (RNP) delivery to treat CHB. In addition, we discuss the importance of delivery tools in the enhancement of the antiviral efficacy of CRISPR/Cas9 against HBV infection.

Role of steroid growth promoter on growth performance and meat quality traits in broiler.

Growth promoters are added with broiler feed to boost the overall feed efficiency and growth rate. The current study investigated the effect of dexamethasone (DEX)-a commonly used growth promoter-on the broiler growth rate, meat quality, and muscle biology. Four homogenous groups (20 chicks/group) of broiler one-day-old chicks were fed commercial broiler feed where the treatment groups received 3, 5, and 7 mg/kg of DEX with their diet for 28 d. Feed consumption and body weight were monitored on a daily basis. Muscle samples were collected on 7, 14, 21, and 28 d of the experiment to investigate meat quality and muscular biology. The residue of DEX in meat was detected using thin-layer chromatography. We observed that DEX had substantially decreased (P < 0.05) feed intake, feed efficiency, and overall weight gain in the broiler. While the weight of breast and thigh meat was decreased, the relative meat weight (meat/body weight) was increased significantly in chicks fed DEX. Simultaneously, body fat decreased while the percentage of fat increased significantly (P < 0.05) in the DEX groups. Contrariwise, DEX improved the investigated meat quality parameters with the potential threat of accumulation of DEX residue in the meat at a high dose (7 mg/kg). We also observed that DEX significantly increased the number of myofibers and decreased the cross-sectional area of myofibers. Based on these findings, we conclude that DEX reduces feed intake, feed efficiency, and growth rate, but might improve meat quality with a potential risk of residual DEX accumulation if fed at a high dose.

Subtype distribution and expression of the koala retrovirus in the Japanese zoo koala population.

We investigated the proviral copies and RNA expression in koala retrovirus (KoRV)-infected koalas. To ascertain any variation in viral load by institution, age, sex, or body condition score, we quantified KoRV proviral DNA and RNA loads in captive koalas (n = 37) reared in Japanese zoos. All koalas were positive for KoRV genes (pol, LTRs, and env of KoRV-A) in genomic DNA (gDNA), and 91.89% were positive for the pol gene in RNA. In contrast, the distribution rates of KoRV-B, KoRV-C, KoRV-D, and KoRV-F env genes in gDNA were 94.59%, 27.03%, 67.57%, and 54.05%, respectively. A wide inter-individual variation and/or a significant inter-institutional difference in proviral DNA (p < 0.0001) and RNA (p < 0.001) amounts (copies/103 koala ß-actin copies) were observed in Awaji Farm England Hill Zoo koalas, which were obtained from southern koala populations, suggesting exogenous incorporation of KoRV in these koalas. Significant (p < 0.05) age differences were noted in KoRV RNA load (p < 0.05) and median total RNA load (p < 0.001), with loads higher in younger koalas (joeys and juveniles). Thus, the current study provides the distribution of KoRV subtypes in Japanese zoo koala populations and identifies several additional risk factors (sex, age, and body condition) associated with KoRV expression.

Bio-intervention phyto-based material for raw goatskin preservation: a cleaner-sustainable approach.

The regular practice of using sodium chloride to preserve raw animal skin triggers increasing salinity and total dissolved solids (TDS) in the surface and groundwater during rehydration soaking operations. The process disrupts the lives of animals, plants, and human beings. This paper is focused on the phyto-based short-term preservation of goatskin to reduce salinity in tannery soaking operations. The indigenous Persicaria hydropiper leaf was investigated to assess the preservation of animal skin to diminish salinity and TDS of tannery soaking wastewater. Methanol-extracted leaf was characterized by GC-MS and FTIR for chemical composition analysis and affiliated functional groups. Fresh goatskins were preserved at the preliminary, laboratory, and pilot-scale scenarios to establish the best possible mixture and monitor the moisture and nitrogen content, shrinkage temperature, microorganism analysis, and pollution load at each level. The processed leathers derived from the preserved skins with an optimal mixture of 10% leaf paste with 8% salt and conventional 50% salt were tested for their physical strength. Finally, the modification in fiber structure due to the varieties of preserving chemicals was evaluated through a scanning electron microscope (SEM) and detected insignificant variation of leather fibers. The findings reported in this study can be applied to the industrial level and remove certain amounts of salinity and TDS from tannery soaking wastewater.

Lead and other elements-based pollution in soil, crops and water near a lead-acid battery recycling factory in Bangladesh.

Lead (Pb) pollution in the environment predominantly occurs through anthropogenic activities, which pose significant threats to human health and that of biota. In this study, Pb and other elements were investigated in different soils (n = 52), crops (n = 24) and water (n = 13) around a lead-acid battery (LAB) recycling workshop in southwestern Bangladesh. Most of the elements' concentrations (except Se and Ag) in soil were lower than the background concentrations. However, excessive concentrations of Pb were found in both surface (966 ± 2414 mg kg-1 at 0-15 cm) and subsurface (230 ± 490 mg kg-1 at 15-30 cm) soil. Although no definitive pattern or direction in elemental concentration in soil was observed, relatively higher concentrations of most elements were detected at the southeast part of the factory. The LAB factory, brick kiln, agricultural and geogenic activities might be the sources of these elements in soil. Extremely high amounts of Cr, As, Cd, and Pb were found in the food crops around the area. In particular, the Pb concentrations were 114 ± 155 and 665 ± 588 mg kg -1 dry weight in rice grain and straw, respectively, which reflected the emissions of Pb from the LAB recycling workshop. Moreover, 40% and 100% of the groundwater samples exceeded, respectively, the WHO provisional guideline values for As (0.01 mg L-1) and Pb (0.05 mg L-1). Consequently, a high level of Pb contamination in the soil was observed while assessing different soil pollution indices. Human health risk assessment indicated severe carcinogenic (from Pb, As, and Cr intake) and non-carcinogenic (from Pb, As, Co, Cr, Ni and Sb intake) health risks are associated with rice and groundwater consumption. It is concluded that all LAB recycling workshops should be better managed to prevent Pb pollution from seeping into the environment.

The environmental impacts of heavy metals in soil, certain plants and wastewater near industrial area of Brahmanbaria, Bangladesh.

At various locations around the world, environments are now becoming greatly polluted by heavy metals, while damage is being done to soil and aquatic life with serious implications for humanity as well. Once heavy metals enter the ecosystem, they persist for a long time, and they are very difficult to eliminate. Discharging of industrial wastes enriched with heavy metals substantially pollutes the soil, water, and air. This study investigated the levels of heavy metals around the industrial area of Brahmanbaria, which is located in Chittagong Division, Bangladesh. Samples of soil, wastewater, vegetable, and grass were collected from the industrial area. The heavy metals were quantified using atomic absorption spectroscopy. The heavy metals found in soil were chromium (Cr) 8.2-18.8, lead (Pb) 3.5-18.3, copper (Cu) 4.6-10.8, zinc (Zn) 48.5-273.4, nickel (Ni) 10.06-26.9, and cadmium (Cd) 0.2 mg/kg, respectively. The metal contents in the wastewater were Cr 0.03-0.5, Pb 0.01-0.05, Cu 0.01-0.02, Zn 0.6-11.2, Ni 0.03-0.14, and Cd 0.003 mg/L, respectively. The metal contents in vegetable and grass were Cr 81.5-247.8, Pb 10.3-34.1, Cu 5.5-6.3, Zn 419.8-435.3, and Ni 8.7-15.5 mg/kg, respectively. The amount of metal in the soil, wastewater, and vegetables and grass followed the sequences: Zn > Cr > Ni > Pb > Cu > Cd, Zn > Cr > Ni > Pb > Cu > Cd, and Zn > Cr > Pb > Ni > Cu, respectively. Compared to the WHO guidelines, the soil in the study area was not polluted with Pb, Cu, and Cd but was moderately polluted with Cr, Zn, and Ni. Continual accumulation of metals in the soil, vegetables and grass could pose a serious risk to the environment as well as many life forms.

Toll-Like Receptor and Cytokine Responses to Infection with Endogenous and Exogenous Koala Retrovirus, and Vaccination as a Control Strategy.

Koala populations are currently declining and under threat from koala retrovirus (KoRV) infection both in the wild and in captivity. KoRV is assumed to cause immunosuppression and neoplastic diseases, favoring chlamydiosis in koalas. Currently, 10 KoRV subtypes have been identified, including an endogenous subtype (KoRV-A) and nine exogenous subtypes (KoRV-B to KoRV-J). The host's immune response acts as a safeguard against pathogens. Therefore, a proper understanding of the immune response mechanisms against infection is of great importance for the host's survival, as well as for the development of therapeutic and prophylactic interventions. A vaccine is an important protective as well as being a therapeutic tool against infectious disease, and several studies have shown promise for the development of an effective vaccine against KoRV. Moreover, CRISPR/Cas9-based genome editing has opened a new window for gene therapy, and it appears to be a potential therapeutic tool in many viral infections, which could also be investigated for the treatment of KoRV infection. Here, we discuss the recent advances made in the understanding of the immune response in KoRV infection, as well as the progress towards vaccine development against KoRV infection in koalas.

Manganese in potable water of nine districts, Bangladesh: human health risk.

Safe drinking water is directly linked to good human health. An excessive amount of manganese (Mn) in drinking water supplies causes people show symptoms of neurotoxicity. In this study, the level of Mn in potable water sourced from tube wells located in 9 (nine) districts of Bangladesh was monitored. In total, 170 (one hundred and seventy) water samples were collected and Mn was quantified by atomic absorption spectroscopy (AAS). The levels of Mn found in the tube well water samples of Sirajganj, Meherpur, Chuadanga, Jhenaidah, Magura, Faridpur, Jashore, Satkhira, and Khulna were 0.37-1.86, 0.10-4.11, 0.30-0.76, 0.26-0.94, 0.01-0.18, 0.21-1.78, 0.08-1.23, 0.05-0.27, and 0.01-2.11 mg/L, respectively. Results revealed that Mn level was beyond the highest contaminated levels of 0.1 mg/L and 0.4 mg/L, which are recommended by Bangladesh Drinking Standard (BDS) and World Health Organization (WHO), respectively. The maximum Mn contaminated level reached up to 4.11 mg/L (mean, 0.53 mg/L). The Mn level in tube well water exceeded 51.1% and 75.9% set by the recommended value of WHO and BDS, respectively. Furthermore, the calculated hazard quotient (HQ) value for Mn was observed to be greater than unity, indicating both children and adults risked potential non-carcinogenic health issues. The water supply authorities should take steps to provide Mn-free drinking water for communities.

Toll-Like Receptor Expression Profiles in Koala (Phascolarctos cinereus) Peripheral Blood Mononuclear Cells Infected with Multiple KoRV Subtypes.

Toll-like receptors (TLRs), evolutionarily conserved pattern recognition receptors, play an important role in innate immunity by recognizing microbial pathogen-associated molecular patterns. Koala retrovirus (KoRV), a major koala pathogen, exists in both endogenous (KoRV-A) and exogenous forms (KoRV-B to J). However, the expression profile of TLRs in koalas infected with KoRV-A and other subtypes is yet to characterize. Here, we investigated TLR expression profiles in koalas with a range of subtype infection profiles (KoRV-A only vs. KoRV-A with KoRV-B and/or -C). To this end, we cloned partial sequences for TLRs (TLR2-10 and TLR13), developed real-time PCR assays, and determined TLRs mRNA expression patterns in koala PBMCs and/or tissues. All the reported TLRs for koala were expressed in PBMCs, and variations in TLR expression were observed in koalas infected with exogenous subtypes (KoRV-B and KoRV-C) compared to the endogenous subtype (KoRV-A) only, which indicates the implications of TLRs in KoRV infection. TLRs were also found to be differentially expressed in koala tissues. This is the first report of TLR expression profiles in koala, which provides insights into koala's immune response to KoRV infection that could be utilized for the future exploitation of TLR modulators in the maintenance of koala health.

Koala retrovirus (KoRV) subtypes and their impact on captive koala (Phascolarctos cinereus) health.

Koala retrovirus (KoRV), a major pathogen of koalas, exists in both endogenous (KoRV-A) and exogenous forms (KoRV-B to J). However, the impact of infection with multiple subtypes is not well understood. Accordingly, in this study, we surveyed a representative sample from a Japanese zoo population to determine the infection status for three KoRV subtypes (KoRV-A, B, and C) and to investigate the proviral and RNA load profiles in animals with single- and multiple-subtype infections, using peripheral blood mononuclear cells (PBMCs) and plasma. Six koalas were evaluated in the study; all were infected with KoRV-A, and two koalas were coinfected with non-A subtypes (KoRV-B and/or KoRV-C). The highest KoRV total RNA and viral loads in PBMCs and plasma were found in a koala infected with multiple subtypes (KoRV-A, -B and -C). The other koala infected with multiple subtypes (KoRV-A and B) showed the highest proviral PBMC load but the lowest RNA copy number in PBMC and plasma. PBMCs from this animal were cultured for further investigation, and KoRV RNA was detected in the cells and culture supernatant after 7 and/or 14 days. The koalas harboring multiple subtypes had a higher white blood cell count than those harboring only KoRV-A and were judged to be leukemic, and they subsequently died due to lymphoma. Accordingly, we conclude that coinfection with multiple KoRV subtypes may be linked to more-severe disease. In a sequence alignment, the detected KoRV-A env gene showed 100% sequence identity to the reference gene, whereas the KoRV-B and -C env genes varied from their reference sequences.

Effects of estradiol on in vitro maturation of buffalo and goat oocytes.

PURPOSE: The effects of estradiol on oocyte development seem to be varied among species. The present study investigated the effects of 17ß-estradiol on in vitro maturation of buffalo and goat oocytes. METHODS: Cumulus oocyte complexes (COCs) were aspirated from large antral follicles of slaughtered buffalo and goat ovaries. COCs were cultured in TCM-199 medium supplemented with 0, 0.5, 1, and 1.5 µg/mL of 17ß-estradiol for in vitro maturation. Then, oocytes were used for the examination of state of nuclear maturation and cumulus expansion. RESULTS: In both species, oocytes treated with 17ß-estradiol showed higher cumulus expansion rate than control (0 µg/mL treated). In buffalo, the percentage of oocytes matured to the metaphase II (MII) stage increased in the concentration-dependent manner of 17ß-estradiol. Similarly, estradiol positively influenced nuclear maturation of goat oocytes in vitro. CONCLUSIONS: Estradiol has promoting effects on normalprogress of in vitro oocyte meiosis in buffalos and goats.

Goatskin preservation with plant oil: significant chloride reduction in tannery wastewater.

Preservation of raw hide/skin is the prime technique to stop bacterial deterioration. Universally, salt curing is the most prevalent technique for hide/skin preservation. In this study, extracted oil from Aphanamixis polystachya seed was evaluated for the preservation of goatskin to reduce the chloride in tannery wastewater. The oil-induced goatskin preservation method was assessed observing diverse factors, e.g., hair slip, odor, moisture content, bacterial colony counting, total Kjeldahl nitrogen, and hydrothermal stability in evaluation to the conventional salt curing method. Results indicate that 15% oil-induced preservation technique could preserve the goatskin for 30 days. A small-scale experiment was performed for the preservation of goatskins. The preserved goatskins both monitoring and study samples were processed for shoe upper leather. The leather quality was evaluated by examining physical properties and fiber structure by a scanning electron microscope (SEM). Pollution load generated during leather making was determined in terms of total dissolved solids (TDS), chloride (Cl-), chemical oxygen demand (COD), and biochemical oxygen demand (BOD). The proposed oil-induced preservation method reduced Cl-, TDS, and BOD by 98.3%, 82.3%, and 86.8%, respectively, in the soaking liquor. The leather readied from the empirical goatskin shows the equipotential properties of leather from conventional goatskin. The oil-induced preservation method is substantiated to be favorable nonconventional preservation instead of conventional wet-salting for the contraction of pollution from soaking operation.

Metagenomic identification, sequencing, and genome analysis of porcine hepe-astroviruses (bastroviruses) in porcine feces in Japan.

Recently, hepe-astrovirus-like RNA viruses named bastroviruses (BastVs), have been found in human, pig, bat, and rat fecal samples. In this study, we determined nearly complete genome sequences of four BastVs in the feces of healthy pigs. Genetic characterization revealed that these porcine BastVs (PBastVs) and BastVs from other animals including humans, had the same genome organization, that is, they contained three predicted conserved domains of viral methyltransferase, RNA helicase, and RdRp in the nonstructural ORF1 and the astrovirus capsid domain in the structural ORF2. Phylogenetic analyses using RNA-dependent RNA polymerase and the capsid region revealed that PBastVs branched with bat and rat BastVs; however, the groups formed by each host were distantly related to human BastVs. Pairwise amino acid sequence comparison demonstrated that PBastVs shared 95.2-98.6% and 76.1-95.5% sequence identity among each other in the ORF1 and ORF2 regions, respectively; the sequence identities between PBastVs and BastVs from other animals were 21.4-42.5% and 9.1-20.6% in the ORF1 and ORF2 regions, respectively. This suggested that BastVs were derived from a common ancestor but evolved independently in each host population during a prolonged period. Putative recombination events were identified in the PBastV genome, suggesting that PBastVs gain sequence diversity and flexibility through recombination events. In an analysis of previously obtained metagenomic data, PBastV sequence reads were detected in 7.3% (23/315) of fecal samples from pigs indicating that PBastVs are distributed among pig populations in Japan.