Phenotypic and genotypic characterization of colistin-resistant Escherichia Coli with mcr-4, mcr-5, mcr-6, and mcr-9 genes from broiler chicken and farm environment.

BACKGROUND: Colistin is an antibiotic used as a last-resort to treat multidrug-resistant Gram-negative bacterial infections. Colistin had been used for a long time in veterinary medicine for disease control and as a growth promoter in food-producing animals. This excessive use of colistin in food animals causes an increase in colistin resistance. This study aimed to determine molecular characteristics of colistin-resistant Escherichia coli in broiler chicken and chicken farm environments. RESULTS: Four hundred fifty-three cloacal and farm environment samples were collected from six different commercial chicken farms in Kelantan, Malaysia. E. coli was isolated using standard bacteriological methods, and the isolates were tested for antimicrobial susceptibility using disc diffusion and colistin minimum inhibitory concentration (MIC) by broth microdilution. Multiplex PCR was used to detect mcr genes, and DNA sequencing was used to confirm the resistance genes. Virulence gene detection, phylogroup, and multilocus sequence typing (MLST) were done to further characterize the E. coli isolates. Out of the 425 (94%; 425/453) E. coli isolated from the chicken and farm environment samples, 10.8% (48/425) isolates were carrying one or more colistin-resistance encoding genes. Of the 48 colistin-resistant isolates, 54.2% (26/48) of the mcr positive isolates were genotypically and phenotypically resistant to colistin with MIC of colistin ≥ 4 µg/ml. The most prominent mcr gene detected was mcr-1 (47.9%; 23/48), followed by mcr-8 (18.8%; 9/48), mcr-7 (14.5%; 7/48), mcr-6 (12.5%; 6/48), mcr-4 (2.1%; 1/48), mcr-5 (2.1%; 1/48), and mcr-9 (2.1%; 1/48) genes. One E. coli isolate originating from the fecal sample was found to harbor both mcr-4 and mcr-6 genes and another isolate from the drinking water sample was carrying mcr-1 and mcr-8 genes. The majority of the mcr positive isolates were categorized under phylogroup A followed by phylogroup B1. The most prevalent sequence typing (ST) was ST1771 (n = 4) followed by ST206 (n = 3). 100% of the mcr positive E. coli isolates were multidrug resistant. The most frequently detected virulence genes among mcr positive E. coli isolates were ast (38%; 18/48) followed by iss (23%; 11/48). This is the first research to report the prevalence of mcr-4, mcr-5, mcr-6, mcr-7, and mcr-8 genes in E. coli from broiler chickens and farm environments in Malaysia. CONCLUSION: Our findings suggest that broiler chickens and broiler farm environments could be reservoirs of colistin-resistant E. coli, posing a risk to public health and food safety.

Evaluation of anti-hyperuricemic effects of Alocasia longiloba Miq. (Keladi Candik) extracts in potassium oxonate induced rat model.

Hyperuricemia has become a significant public-health concern in recent years, and the available treatments have been reported to have an adverse side effect on patients. Alocasia longiloba has been used traditionally in Malaysia for treating gout, inflammation, and wounds. However, the plant has not been investigated for its effects on hyperuricemia. This study investigated the anti-hyperuricemic and anti-inflammatory effects of A. longiloba extracts in hyperuricemic rats induced by potassium oxonate (250 mg/kg body weight). Rats were given A. longiloba extracts or a standard drug for two-week, and blood and tissue samples were collected for analysis. Results show that A. longiloba extracts significantly reduced serum uric acid levels in hyperuricemic rats and inhibited xanthine oxidase (XOD) activity in the liver and kidney, which could be the mechanism underlying the urate-lowering effects. The extracts also significantly (p < 0.05) reduced the levels of proinflammatory cytokines (IL-18 and IL-1ß) in serum samples and had hepatoprotective and nephroprotective effects in hyperuricemic rats. The study supports the use of A. longiloba as a complementary therapy for treating hyperuricemia.

Natural Polymeric Composites Derived from Animals, Plants, and Microbes for Vaccine Delivery and Adjuvant Applications: A Review.

A key element in ensuring successful immunization is the efficient delivery of vaccines. However, poor immunogenicity and adverse inflammatory immunogenic reactions make the establishment of an efficient vaccine delivery method a challenging task. The delivery of vaccines has been performed via a variety of delivery methods, including natural-polymer-based carriers that are relatively biocompatible and have low toxicity. The incorporation of adjuvants or antigens into biomaterial-based immunizations has demonstrated better immune response than formulations that just contain the antigen. This system may enable antigen-mediated immunogenicity and shelter and transport the cargo vaccine or antigen to the appropriate target organ. In this regard, this work reviews the recent applications of natural polymer composites from different sources, such as animals, plants, and microbes, in vaccine delivery systems.

The Roles of Polysaccharides in Carp Farming: A Review.

Carp is an important aquaculture species globally, and the production is expected to increase with the growing market demands. Despite that, disease outbreaks remain a major challenge, impeding the development of sustainable carp farming. Moreover, the application of antibiotics, a common prophylactic agent, can adversely impact public health and the environment. Therefore, polysaccharide has been recognized as a novel prophylactic agent in the health management of carp farming, as well as gaining consumers' confidence in carp farming products. In this review, the definition, sources, and main roles of polysaccharides in improving growth performance, stimulating the immune system, enhancing disease resistance, and alleviating abiotic stresses in carp farming are discussed and summarized. In addition, the use of polysaccharides in combination with other prophylactic agents to improve carp farming production is also highlighted. This review aims to highlight the roles of polysaccharides and provide valuable information on the benefits of polysaccharides in carp farming.

LC-TOF-MS/MS and GC-MS based phytochemical profiling and evaluation of wound healing activity of Oroxylum Indicum (L.) Kurz (Beka).

Background: Beka (Oroxylum indicum (L.) Kurz) has been used as a culinary herb and natural remedy by the local communities in Malaysia. The leaf of O. indicum is traditionally used for the treatment of diarrhea, high blood pressure, and improving digestive health. Objectives: The present study was conducted to evaluate the phytochemical constituents and wound healing properties (in vitro and in vivo models) of aqueous and ethanol extracts of O. indicum leaves. Methods: The total phenolic (TPC) and total flavonoid (TFC) contents in the plant extracts were determined by the spectrophotometric methods. Further, the extract was characterized by Liquid Chromatography Time-of-Flight Mass Spectrometry (LC-TOF-MS/MS) and Gas Chromatography-Mass Spectrometry (GC-MS). The wound healing activity was assessed using the in vitro scratch wound-healing assay and in vivo excisional wound model. Results: The results show the ethanol leaves extract had the higher TPC (164 mg GAE/g) when compared with the aqueous leaves extract (30 mg gallic acid equivalents/g). The ethanol leaves extract was also found to have higher TFC (101 mg Catechin equivalents/g) than the aqueous leaves extract (76 mg Catechin equivalents/g). The ethanol leaves extract was then used for further chemical analysis. The LC-TOF-MS/MS analysis showed that the leaves extracts of O. indicum contains many important compounds such as Orientin, Chrysin, Pinoquercetin, Cupressuflavone, Puerarin xyloside, Forsythiaside and Paederoside. In GC-MS analysis, 19 compounds were identified in ethanolic leaves extract. The wound healing studies shows that O. indicum has promising wound healing activity by increasing the rate of wound contraction significantly (p < 0.05). Conclusion: In conclusion, the present study showed that O. indicum leaf contains important phytochemicals and the wound healing potential of the O. indicum extract may probably be as a result of the presence of various phytoconstituents.

Molecular detection of pathogenic Leptospira spp. in urban rodents from wet markets in northeast Malaysia.

Objective: This short study describes the occurrence of pathogenic Leptospira spp. in two major wet markets in Kota Bharu, Kelantan, Malaysia. Materials and Methods: 30 rodents (20 rats and 10 shrews) were caught in 2 wet markets, and a postmortem was performed to extract both kidneys. Molecular diagnosis via polymerase chain reaction (PCR) was conducted to detect leptospiral DNA using universal and pathogenic Leptospira primers, respectively. Results: The results showed that 20/28 (72%) rat samples were detected positive for Leptospira spp, and all shrews were negative. Further sequencing analysis identified L. interrogans and L. borgpetersenii as the most frequently Leptospirosis species from kidney samples. Conclusions: The presented study here sheds light on the presence of pathogenic leptospires harboring the rat population in both wet markets in Kelantan, which presents a great public health risk to wet market workers and visitors.

A Rare Case of Oral Papillomatosis in a Goat Kid.

The clinical management and outcome of a case of caprine papillomatosis in a 2-week-old kid goat was reported. Based on the PCR result, contagious ecthyma (CE) was ruled out. Based on the history and physical examination findings, the case was diagnosed as severe clinical case of papillomatosis in a goat's kid. The treatment procedure was administrated with flunixin meglumine (5%) 1.1 mg/kg, 0.5 ml, IM, SID, 3 days, and Penstrep (procaine penicillin, dihydrostreptomycin sulphate) 2 ml and a topical preparation for 5 days to prevent secondary bacterial infection. Also, Vitavet (multivitamin-vitamins A, D3, and B12), 1 ml/10 kg, 1 ml, IM, SID, was administered for 3 days to boost the immune system.

First case report on molecular detection of Trypanosoma lewisi in an urban rat in Kelantan, Malaysia: An accidental finding.

OBJECTIVE: This case report highlights the first detection of Trypanosoma lewisi, a blood protozoan parasite found in an urban rat in Kota Bharu, Kelantan. MATERIALS AND METHODS: Rat trapping was carried out within the Kota Bharu vicinity near a local wet market. A total of 38 rats were captured and subjected to peripheral blood smearing using Giemsa stain. Positive rats were sent for histopathological analysis for the evaluation of the organ samples. RESULTS: The presence of trypanosomes was found in one sample from a blood smear. This was connected to a histological lesion on kidney tissues, which revealed a high concentration of trypanosomes. Additionally, the positive sample was confirmed as T. lewisi based on molecular diagnosis via polymerase chain reaction and subsequent sequencing and phylogenetic analysis. CONCLUSIONS: This finding serves as a baseline for further surveillance on T. lewisi population among urban rats in Kelantan and possible zoonotic transmission to humans.

Optimization of Ultrasound-Assisted Extraction of Bioactive Compounds from Acacia Seyal Gum Using Response Surface Methodology and Their Chemical Content Identification by Raman, FTIR, and GC-TOFMS.

Acacia Seyal gum (ASG), also known as gum Arabic, is an antioxidant-rich soluble fiber. ASG has been reported to have many biological activities, including anticancer, antidiabetic, antiulcer, and immunomodulatory activity. Extraction of bioactive compounds from ASG is commonly performed using conventional extraction methods. However, these techniques have certain limitation in terms of extraction time, energy, and solvent requirements. Ultrasound-assisted extraction (UAE) could be used as an alternative technique to extract bioactive compounds in less time, at low temperature, and with less energy and solvent requirements. In this study, the UAE extraction of ASG was optimized using response surface methodology (RSM). A face-centered central composite design (FCCCD) was used to monitor the effect of different independent factors of ultrasound operation (sonication time, temperature, and solvent ratio) on ASG extraction yield. In addition, screening and characterization of phytochemicals in 60% ethanol ASG extract was carried out using Raman microscopy, Fourier transform infrared spectroscopy (FTIR), and gas chromatography time-of-flight mass spectroscopy (GC-TOFMS) analysis. The results indicated that, under optimal conditions (extraction time 45 min, extraction temperature 40 °C, and solid-liquid ratio of 1:25 g/mL), the yield of ASG was 75.87% ± 0.10. This yield was reasonably close to the predicted yield of 75.39% suggested by the design of experiment. The ANOVA revealed that the model was highly significant due to the low probability value (p < 0.0001). Raman spectrum fingerprint detected polysaccharides, such as galactose and glucose, and protein like lysine and proline, while FTIR spectrum revealed the presence of functional groups peaks value of alkanes, aldehydes, aliphatic amines, and phenol. GC-TOFMS spectroscopic detected the presence of strong d-galactopyranose, carotenoid, and lycopene antioxidant compounds. In conclusion, this study demonstrated that the UAE technique is an efficient method to achieve a high yield of ASG extracts. The selected model is adequate to optimize the extraction of several chemical compounds reported in this study.

Rhipicephalus Tick: A Contextual Review for Southeast Asia.

Rhipicephalus species are distributed globally with a notifiable presence in Southeast Asia (SEA) within animal and human populations. The Rhipicephalus species are highly adaptive and have established successful coexistence within human dwellings and are known to be active all year round, predominantly in tropical and subtropical climates existing in SEA. In this review, the morphological characteristics, epidemiology, and epizootiology of Rhipicephalus tick species found in SEA are reviewed. There are six commonly reported Rhipicephalus ticks in the SEA region. Their interactions with their host species that range from cattle, sheep, and goats, through cats and dogs, to rodents and man are discussed in this article. Rhipicephalus-borne pathogens, including Anaplasma species, Ehrlichia species, Babesia species, and Theileria species, have been highlighted as are relevant to the region in review. Pathogens transmitted from Rhipicepahalus ticks to host animals are usually presented clinically with signs of anemia, jaundice, and other signs of hemolytic changes. Rhipicephalus ticks infestation also account for ectoparasitic nuisance in man and animals. These issues are discussed with specific interest to the SEA countries highlighting peculiarities of the region in the epidemiology of Rhipicephalus species and attendant pathogens therein. This paper also discusses the current general control strategies for ticks in SEA proffering measures required for increased documentation. The potential risks associated with rampant and improper acaricide use are highlighted. Furthermore, such practices lead to acaricide resistance among Rhipicephalus species are highlighted.

The influence of feeding low and high level of Brachiaria decumbens diets on the hematology, serum biochemistry, and acute phase proteins of sheep.

The present study aims to determine the hematology, serum biochemistry, and acute phase proteins (APPs) responses of both serum and cerebrospinal fluid (CSF) in sheep fed with low and high levels of Brachiaria decumbens (B. decumbens) diets at different time phases. A total of 30 6-month-old male Dorper cross sheep were randomly divided into three treatment groups consisted of 10 sheep each. Treatment 1 (control) sheep were fed with Pennisetum purpureum and concentrates as the basal diet, whereas Treatments 2 and 3 sheep were fed with low (10%) and high (60%) level of B. decumbens, respectively. The hematology results revealed that there were significant differences (p < 0.05) in the red blood cells, mean corpuscular volume, mean corpuscular hemoglobin concentration, white blood cells, neutrophils, monocytes, eosinophils, basophils, platelets, and plasma proteins between groups. Except for packed cell volume, there were also significant differences in all hematology parameters at different time phases. All biochemistry parameters except creatinine revealed significant differences among treatment groups. However, there were significant differences in all parameters between time. On the other hand, APPs results showed significant differences in the serum haptoglobin and serum amyloid A in both serum and CSF between groups and time.

Infectious bronchitis associated with Escherichia coli infection in commercial broiler chickens: a case report.

OBJECTIVE: In this case report, we have investigated the infectious bronchitis (IB) virus (IBV) outbreak with the co-infection of Escherichia coli in 28-33-day-old broiler chickens in Malaysia. MATERIALS AND METHODS: A farmer complained that Cobb 500 chickens, raised in the open house, were having bloody diarrhea, open mouth breathing, non-uniform growth, and ruffled feathers. The mortality was about 100 birds (from about 7000 birds) per day. The sick birds were isolated and subjected to physical examination, postmortem, and histopathological analyses. Gross lesions were observed and recorded. The lung samples have proceeded with histopathological evaluations. The lungs, kidneys, trachea, air sac, and heart samples were collected to isolate bacteria and fungi through a series of conventional cultural methods, followed by molecular confirmation of the IBV. RESULTS: Postmortem examination revealed air sacculitis, hemorrhagic tracheitis, pulmonary congestion, fibrin deposition in the liver and air sac, hemorrhagic enteritis, and renomegaly. The bacterial culture and biochemical tests revealed E. coli in the lungs, trachea, liver, intestine, and kidney samples. However, no fungus could be isolated from those samples. Histological evaluation of lung samples demonstrated infiltration of inflammatory cells in the pulmonary tissues. Apart from this, reverse transcription-polymerase chain reaction confirmed the presence of avian coronavirus responsible for infectious bronchitis (IB). CONCLUSION: The chickens were diagnosed with IB concurrent with E.coli. The chickens exhibited typical nephropathogenic strain of IBV infection, causing high mortality.

Complex chronic respiratory disease concurrent with coccidiosis in broiler chickens in Malaysia: A case report.

OBJECTIVE: The case study describes the cause of an increase in mortality rates among 35-day-old broilers that developed respiratory distress and bloody diarrhea on a farm in Malaysia. MATERIALS AND METHODS: The organ samples were subjected to laboratory testing and postmortem inspection. Escherichia (E.) coli and Mycoplasma (M.) gallisepticum were detected using bacterial isolation and molecular diagnostics using polymerase chain reaction. RESULTS: Chickens with the infection had widespread fibrin buildup in several organs and hemorrhages on the duodenal mucosa. Additional histology and laboratory analysis of organ samples revealed infection with M. gallisepticum, E. coli, and enteric Eimeria spp., all of which are consistent with complex chronic respiratory disease (CCRD) associated with coccidiosis. Tylosin tartrate 20% (w/w) (2.5 gm/l) was prescribed for 1 week along with a combination of the broad-spectrum bacteriostatic drug streptomycin (25 mg/kg) and coccidiostat (2 gm/5 l). CONCLUSION: CCRD and coccidiosis are both infectious diseases that can infect chicken flocks, resulting in production losses and carcass quality degradation. Early disease detection and proper treatment should be provided promptly, and tight farm biosecurity should be implemented to prevent chicken mortality on the farm, as was achieved successfully.

First case report on the occurrence of Trypanosoma evansi in a Siam B Mare in Kelantan, Malaysia.

This is the first case report for the positive Trypanosoma evansi incident in Kelantan, Malaysia confirmed through protozoa detection in a Siam B mare. The horse was presented with complaints of lethargy and inappetence and it collapsed on the day of visit. Normal saline and dextrose solution were administered intravenously, while multivitamins and nerve supplements were given intramuscularly to stabilise the horse before further treatment. Haematological findings showed normocytic hypochromic anaemia and are suggestive of regenerative anaemia. Thin blood smear and examination revealed the presence of Trypanosoma sp., and it was confirmed as T. evansi through molecular identification. The horse was found dead 2 days after and post-mortem was conducted. Histopathology revealed that the horse had developed a neurological form of the disease, besides the detection of the protozoa in heart, spleen and kidney tissue. This first positive Surra case in Kelantan, Malaysia, that is bordering Thailand confirms the increasing concern of transboundary infections. In conclusion, Surra is a potential emerging disease and should be considered as differential diagnosis in horses with pale mucous membrane. This condition is particularly imperative in horses found in these regions as Surra is endemic.

Polycystic kidney disease concurrent with feline parvovirus and bacterial infections in domestic shorthair cat: a case report.

Feline polycystic kidney disease (PKD) is an inherited disorder caused by the mutation of PKD1 gene that eventually lead to the development of chronic kidney disease. The latter condition causes hypertension and eventually progress into congestive heart failure. Feline parvovirus (FPV) is a highly contagious and often fatal disease infecting cats and other members of Felidae. An 8-month-old female domestic shorthair cat was presented with complaint of wound dehiscence a day after ovarian hysterectomy procedure. The wound at the suture site appeared necrotic, purulent with foul smell. The cat was found to have diarrhoea during the fixation of suture breakdown and, later, was tested positive with parvovirus infection. Complete blood count revealed anaemia, neutrophilia, lymphopenia and thrombocytosis. Biochemistry profiles showed hypoproteinaemia and elevated of urea and creatinine. The cat was hospitalised, and symptomatic treatments were given. During hospitalisation, the cat showed symptoms of polydipsia and polyuria and found dead 2 days later. Post-mortem findings demonstrated the cat had oral ulceration, thoracic effusion, fibrinopleuropneumonia, pericardial effusion, left ventricular hypertrophy and right ventricular dilation, chronic passive liver congestion, mesenteric lymphadenomegaly, intestinal haemorrhage, adrenomegaly and polycystic kidney. Histopathological evaluation revealed fibrinous pleuropneumonia, pulmonary atelectasis, emphysema and oedema, hypertrophic cardiomyopathy, hepatic necrosis, splenic necrosis, intestinal necrosis, renal necrosis and renal polycystic. Staphylococcus aureus and Escherichia coli were isolated from bronchus swab and intestinal segment, respectively. Polymerase chain reaction (PCR) revealed parvovirus infection. The cat was definitely diagnosed with polycystic kidney disease concurrent with parvoviral and secondary bacterial infections.

Micropropagation of Alocasia longiloba Miq and Comparative Antioxidant Properties of Ethanolic Extracts of the Field-Grown Plant, In Vitro Propagated and In Vitro-Derived Callus.

In this study, an efficient micropropagation protocol was developed for A. longiloba and the antioxidant properties of field-grown plant, in vitro-derived greenhouse-grown plant and in vitro-derived callus extracts were compared. The A. longiloba seeds tested using tetrazolium chloride salt exhibited 89% viability. Due to poor germination capacity of A. longiloba seeds, the seeds were treated with gibberellic acid (GA3) or sulfuric acid (H2SO4). The maximum seed germination of 87% was observed at 30% H2SO4 treatment after 19.00 d, whereas GA3 treatment showed maximum germination of 53% after 22 d. In vitro shoot multiplication was carried out using various types of cytokinins alone or in combination with auxin. Among them, 6-benzyl amino purine (BAP) single treatment was found to be the best hormone. The highest shoot-length (7.26 cm) and maximum number of shoots per explant (18) were recorded at 3-mg L-1 BAP. For in vitro rooting, indole-3-acetic acid at 0.5-mg L-1 was found to be the optimum concentration. Callus was induced using various types of auxins alone or in combinations with cytokinins. The highest percentage of callus of 91 and fresh weight of 6 g was obtained with 3-mg L-1 IAA. The plantlets produced in the current study were subjected to acclimatization. The combination of topsoil and peat moss at 1:2 ratio was found to be the best soil media. In this study, in vitro-derived callus extract showed the highest phenolic content (538 mg GAE), followed by extracts of field-grown plant parts, i.e., fruit and petiole (504 and 300 mg GAE) while in vitro plant extract showed the lowest (98 mg GAE). Meanwhile, the highest flavonoids was recorded in petiole extract. Comparative antioxidant activity study shows, in vitro-derived callus exhibited better DPPH-radical-scavenging activity (IC50: 0.113-mg mL-1) whereas the extracts of petiole, fruit and in vitro plant showed 0.126-, 0.137- and 0.173-mg mL-1, respectively. At the same time, the fruit extract showed better (IC50: 0.088-mg mL-1) ABTS radical scavenging activity than all extracts tested. In conclusion, the in vitro-derived callus extract could be favored for high TPC and better DPPH scavenging activity. Hence, the present study was conducted to establish an efficient micropropagation protocol and to compare the antioxidant activity of the field-grown plant, in vitro plant and in vitro derived callus extracts of A. longiloba.

Subacute Oral Administration of Clinacanthus nutans Ethanolic Leaf Extract Induced Liver and Kidney Toxicities in ICR Mice.

This study investigated the leaves of Clinacanthus nutans for its bioactive compounds and acute and subacute toxicity effects of C. nutans ethanolic leaf extract (CELE) on blood, liver and kidneys of ICR mice. A total of 10 8-week-old female mice were divided into groups A (control) and B (2000 mg/kg) for the acute toxicity study. A single dose of 2000 mg/kg was administered to group B through oral gavage and mice were monitored for 14 days. In the subacute toxicity study, mice were divided into five groups: A (control), B (125 mg/kg), C (250 mg/kg), D (500 mg/kg) and E (1000 mg/kg). The extract was administered daily for 28 days via oral gavage. The mice were sacrificed, and samples were collected for analyses. Myricetin, orientin, isoorientin, vitexin, isovitexin, isookanin, apigenin and ferulic acid were identified in the extract. Twenty-eight days of continuous oral administration revealed significant increases (p < 0.05) in creatinine, ALT and moderate hepatic and renal necrosis in groups D and E. The study concluded that the lethal dose (LD50) of CELE in mice is greater than 2000 mg/kg and that repeated oral administrations of CELE for 28 days induced hepatic and renal toxicities at 1000 mg/kg in female ICR mice.

N-Ethyl-n-Nitrosourea Induced Leukaemia in a Mouse Model through Upregulation of Vascular Endothelial Growth Factor and Evading Apoptosis.

Chemical carcinogens are commonly used to investigate the biology and prognoses of various cancers. This study investigated the mechanism of leukaemogenic effects of n-ethyl-n-nitrosourea (ENU) in a mouse model. A total of 14 3-week-old male Institute of Cancer Research (ICR)-mice were used for the study. The mice were divided into groups A and B with seven mice each. Group A served as the control while group B received intraperitoneal (IP) injections of 80 mg/kg ENU twice with a one-week interval and were monitored monthly for 3 months for the development of leukaemia via blood smear examination. The mice were sacrificed humanely using a CO2 chamber. Blood, spleen, lymph nodes, liver, kidney and lung samples were collected for blood smear examination and histopathological evaluation. The expression of angiogenic protein (VEGF), and pro and anti-apoptotic proteins (BCL2 and BAX), was detected and quantified using Western blot technique. Leukaemia was confirmed by the presence of numerous blast cells in the peripheral blood smear in group B. Similarly, the VEGF and BCL2 proteins were significantly (p < 0.05) upregulated in group B compared to A. It is concluded that IP administration of 80 mg/kg ENU induced leukaemia in ICR-mice 12 weeks post administration through upregulation of angiogenic and anti-apoptotic proteins: VEGF and BCL2.

A case of aspergillosis outbreak in a broiler duck farm in Kelantan, Malaysia.

OBJECTIVE: This case report aims to discuss the veterinary approach taken to control a case of aspergillosis outbreak on a duck farm. MATERIALS AND METHODS: A broiler duck farm with a population of 900 Muscovy ducks was having a complaint of a 5% mortality rate in their 3-week-old ducklings. Upon presentation, 10% of the ducks appeared to be listless, dyspneic, ruffled feathers, and cyanotic. Postmortem examination of the dead birds was conducted. The collected samples were subjected to isolation and identification of the associated Aspergillus fumigatus under the microscope using the scotch tape method. RESULTS: Postmortem examination revealed whitish to creamy caseous nodules in the lungs, thoracic air sacs, gizzard, proventriculus, and intestines. Granuloma lesions and infiltration of inflammatory cells were observed in the lung and liver tissues. As for therapeutic management, all ducks were treated with copper sulfate, erythromycin, and multivitamins as the fungicide, antibiotic, and supplement, respectively, via drinking water. CONCLUSION: There is no effective treatment for Aspergillosis as the spores are difficult to destroy completely. Nonetheless, the disease can be controlled and prevented effectively with proper farm sanitation and providing a suitable feed storage environment to inhibit the growth of this opportunistic fungus.