The Optimal Dosage of Fermented Herbal Extract on Growth and Feed Efficiency of Nile Tilapia (Oreochromis niloticus).

Owing to their availability, cost effectiveness and environmental-friendly nature, plant extracts are promising additives for fish farming. This study aims to determine the optimal dosage of fermented herbal extract (FHE)-composed of Morus alba (33.3%), Curcuma xanthorrhiza (33.3%), and Boesenbergia rotunda (33.3%)-for growth enhancement and feed utilisation efficiency of Oreochromis niloticus fingerlings. Fermentation was conducted using probiotics Lactobacillus casei (Yakult®, Tokyo, Japan) and Saccharomyces cereviceae (commercial baker's yeast). The FHE was high in flavonoid and alkaloid, vitamin C, potassium, natrium, lipase and protease. Four doses of FHE treatments, namely treatment A (0 mg/kg of feed); treatment B (100 mg/kg of feed); treatment C (300 mg/kg of feed); treatment D (500 mg/kg of feed) were compared. After subjected to 35 days of culture, tilapias subjected to FHE-coated feed exhibited better weight gain (WG), specific growth rate (SGR), and feed efficiency (FE) compared to control. The best dosage that gave the highest growth and feed efficiency was treatment C (300 mg/kg of feed). Furthermore, the feed efficiencies of FHE-incorporated treatments were positively influenced by the increased in length and density of intestinal villi, number of goblet cells, lymphocytes, as well as nutrient retention to support growth. The results of this study indicate that FHE is a promising functional feed additive to stimulate growth and improve feed efficiency in tilapia farming.

Phylogenetic analysis of HPAI H5N1 virus from duck swab specimens in Indonesia.

OBJECTIVE: A phylogenetic study was carried out on the avian influenza virus (AIV) isolated from a disease outbreak in Sidenreng Rappang Regency, South Sulawesi, Indonesia, in 2018. MATERIAL AND METHODS: Oropharyngeal swabs and organ samples were obtained from ducks that showed clinical symptoms: torticollis, fascial edema, neurological disorders, the corneas appear cloudy, and death occurs less than 1 day after symptoms appear. In this study, isolate A/duck/Sidenreng Rappang/07180110-11/2018 from duck was sequenced and characterized. RESULTS: It was found that each gene segment of the virus has the highest nucleotide homology to the Indonesian highly pathogenic avian influenza (HPAI) H5N1 clade 2.3.2.1c. Multiple alignments of the sample Hemagglutinin (HA) gene with the avian influenza references virus showed that the pattern of amino acid arrangement in the cleavage site PQRERRRK-RGLF is the characteristic of the HPAI virus. In addition, the HA gene contained Q222 (glutamine) and G224 (glycine), signifying a high affinity to avian receptor binding specificity (SA α2,3 Gal). Furthermore, there was no genetic reassortment of this virus based on the phylogenetic analysis of HA, NA, PB1, PB2, PA, NP, M, and NS genes. CONCLUSION: The HPAI H5N1 clade 2.3.2.1c virus was identified in duck farms in South Sulawesi, Indonesia.

Development of neutralization antibodies against highly pathogenic H5N1 avian influenza virus using ostrich (Struthio camelus) yolk.

The rapid outbreak of the highly pathogenic H5N1 avian influenza virus and its transmission to humans have induced world-wide fears of a new influenza pandemic. The most effective method for the reduction of the impact of such a pandemic would be prophylaxis with a safe and effective vaccine, as well as anti-viral materials. In this study, we generated the specific antibodies 'immunoglobulin yolk (IgY)' from ostrich eggs immunized with a full-length glycosylated recombinant H5 protein of the strain H5N1/Vietnam/1203/2004. Using this simple method, abundant specific antibody (about 200 g) against H5 was successfully produced by one female ostrich in a year. The IgY from the immunized ostrich eggs had strong reactivity to the H5N1 virus as well as to H5 proteins. Furthermore, the antibodies strongly inhibited cytopathic effects in MDCK cells and prevented the death of an embryonated chick after a viral inoculation, indicating strong neutralization activity against H5N1 infections. These findings suggest that the neutralization antibody produced by the H5-immunized ostrich is suitable for industrial purposes, such as the development of antibody-binding filters, which can be applied to a mask or to air-conditioners to prevent the influenza pandemic through antigen-antibody reactions. Of note, the mortality rate of chicks inoculated with the H5N1 virus was dramatically decreased with antibody injection. This indicates that ostrich IgY is a potentially effective therapeutic modality for H5N1 infection.

Expression of calbindin-D28K in sporadic nephroblastomas of the chicken.

Calbindin-D28K (Ca-D28K) is a calcium-binding protein. In the kidney, Ca-D28K is present in the distal nephron, but not in the proximal nephron. This site-specific distribution in the kidney indicates that Ca-D28K is a potential marker for the differentiation of the distal nephron. In this study, we have examined the expression of Ca-D28K in 25 sporadic cases of chicken nephroblastomas. All cases of nephroblastomas were composed of atypical tubular structures, blastemal cells, and fibrous stroma in varying degrees of differentiation. Immunohistochemically in all nephroblastoma specimens, Ca-D28K was expressed in the epithelial cells of the subsets of tubular structures, but not in the blastema or the stroma. These results suggested that the tubuli in the nephroblastomas are able to differentiate into the phenotype of distal nephrons. Furthermore, Ca-D28K might develop as a novel diagnostic marker for nephroblastomas because this molecule is reported to be completely negative in other renal tumors, including renal cell carcinoma, chromophobe carcinomas, and oncocytoma.

Hypothalamo-pituitary-adrenal gland axis in mice inhaling toluene prior to low-level long-term exposure to formaldehyde.

We studied the change in the hypothalamo-pituitary-adrenal gland (HPA) axis upon adding prior toluene inhalation to our previous formaldehyde inhalation experiments to determine whether short term exposure to relatively high levels of toluene triggers multiple chemical sensitivity (MCS). Data come from immunocytochemical, morphometrical and RT-PCR measurements. Four groups of adult female mice were exposed to differing concentrations (0, 80, 400, and 2,000 ppb) of formaldehyde for 16 hr/day, 5 days/week for twelve weeks, after the mice were exposed intranasally to 500 ppm toluene per mouse for 6 hr/day, for 3 days. We found that the number of corticotropin releasing hormone (CRH)-immunoreactive (ir) neurons was up-regulated according to the amount of formaldehyde as well as inhalation of formaldehyde alone in our previous experiment. The proportion of adrenocorticotropin hormone (ACTH)-ir cells increased according to the formaldehyde concentration, though there was no significant difference between the 400 and 2,000 groups. The number of ACTH-ir cells was higher in the 400 group than in the other groups (0, 80, and 2,000). Expression of ACTH-mRNA was also up-regulated according to the quantity of formaldehyde. The sinusoid in the anterior pituitary showed more dilatation in the 400 and 2,000 groups than in the control group, especially in the 2,000 group. We propose that exposure to toluene prior to inhalation of formaldehyde has no effect on the HPA axis and as a trigger of MCS, although greater sinusoid dilatation was found in the anterior pituitary gland at higher concentrations of formaldehyde.

Effect of prolonged exposure to low concentrations of formaldehyde on the corticotropin releasing hormone neurons in the hypothalamus and adrenocorticotropic hormone cells in the pituitary gland in female mice.

We examine the effect on the hypothalamus-pituitary-adrenal gland (HPA) axis of prolonged exposure to low levels of formaldehyde in female C3H/He mice, using immunocytochemical and RT-PCR methods. Two groups of female mice were exposed to differing concentrations (0, 80, 400, 2000 ppb) of formaldehyde inhalation for 16 h/day, 5 days/week, for 12 weeks. The corticotropin releasing hormone (CRH)-immunoreactive (ir) neurons in the hypothalamus were then examined, together with the adrenocorticotropin hormone (ACTH)-ir cells and ACTH mRNA in the pituitary. One group comprised sham control mice. The other group was made allergic by injection of ovalbumin (OVA) and alum prior to exposure to formaldehyde, since most sick building syndrome (SBS) sufferers are women with allergic disease. These animals were further exposed to aerosolized OVA as a booster four times during the exposure period. Our results showed a dose-dependent increase in the number of CRH-ir neurons in the non-allergy (NAG) group. A similar pattern was found in ACTH-ir cells and ACTH mRNA. The allergy (AG) model group showed an increase in basal levels of all markers of HPA activity. Moreover, the AG mice appeared to respond to the lowest concentration of formaldehyde, and all indices of HPA activity were reduced at the highest concentrations of formaldehyde. These results relate to an important clinical issue and also have implications in the broader area of HPA regulation. We conclude that our experimental system may be a suitable animal model for SBS and/or multiple chemical sensitivity (MCS).

Age-related changes in growth hormone (GH) cells in the pituitary gland of male mice are mediated by GH-releasing hormone but not by somatostatin in the hypothalamus.

Using immunocytochemical and morphometric methods, we examine changes with age of growth hormone-releasing hormone (GHRH) in the arcuate nucleus (ARC), changes of somatostatin (SS) in the periventricular nucleus (PeN) of the hypothalamus, and changes of growth hormone (GH) cells in the anterior pituitary in male C57BL/6J mice at 2 months old (2 M), 4 M, 12 M and 24 M. The number of GHRH-ir neurons decreased significantly with age. The number of SS-ir neurons did not differ significantly between these all age groups. The volume of the anterior pituitary and the number of adenohypophysial parenchymal cells fell dramatically from 4 to 12 M. The proportion of GH-ir cells decreased significantly with age, and in absolute number from 4 to 12 M and in size from 2 to 4 M and from 4 to 12 M. These results suggest that the reduction in GH-ir cells in male mice is modulated by the reduction in GHRH-ir neurons, but not by SS-ir neurons.