Classification of Cryptococcus neoformans and yeast-like fungus isolates from pigeon droppings by colony phenotyping and ITS genotyping and their seasonal variations in Korea

Cryptococcus neoformans (C neoformans) is a frequent cause of invasive fungal disease in immunocompromised human hosts. Ninety-eight samples of pigeon droppings were collected from the pigeon shelters in Seoul, and cultured on birdseed agar (BSA) and Sabouraud dextrose agar (SDA). One hundred yeast-like colonies were selected and identified via phenotype characteristics, such as colony morphology and biochemical characteristics. This was then followed with genotyping via sequencing of the internal transcribed spacer (ITS) region. The colonies were classified into four kinds of colony color types: brown type (BrT), beige type (BeT), pink type (PT), and white type (WT). Numbers of isolated BrT, BeT, PT, and WT colonies were 22 (22%), 30 (30%), 19 (19%), and 39 (39%), respectively. All BrT colonies were identified as C neoformans. BeT were identified as 19 isolates of Cryptococcus laurentii, 10 isolates of Malassezia furfur, and 1 isolate of Cryptococcus uniguttulatus. PT was divided into two colony color types: light-PT (l-PT) and deep-PT (d-PT). Eighteen of l-PT and one of d-PT were identified as Rhodotorula glutinis and Rhodotorula mucilaginosa, respectively. WT were identified as 34 isolates of Cryptococcus guilliermondii, 3 isolates of Cryptococcus zeylanoides, 1 isolate of Cryptococcus sake, and 1 isolate of Stephanoascus ciferrii. Most strains were classified identically with the use of either phenotype or genotyping techniques, but C uniguttulatus and C sake classified by phenotyping were Pseudozyma aphidis and Cryptococcus famata by genotyping. This rapid screening technique of pathogenic yeast-like fungi by only colony characteristics is also expected to be very useful for primary yeast screening. Additionally, we investigated the seasonal variations of C neoformans and other yeast-like fungi from 379 pigeon-dropping samples that were collected from February 2011 to March 2011. We isolated 685 yeast-like fungi from the samples. Almost all C neoformans and yeast-like fungi were isolated in the fall (298 strains, 43.5%) and spring (244 strains, 35.6%). A few yeast-like fungi were isolated in winter (98 strains, 14.3%) and summer (45 strains, 6%). These results would be used as an important indicator related to epidemiology and prevention of pathogenic yeast-like fungi infections transmitted through pigeon droppings.     

Development and characterization of mouse monoclonal antibodies reactive with chicken CD83

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD83 (chCD83), a membrane-bound glycoprotein belonging to the immunoglobulin superfamily that is primarily expressed on mature dendritic cells (DCs). A recombinant chCD83/IgG4 fusion protein containing the extracellular region of chCD83 was expressed in Chinese Hamster Ovary (CHO) cells and isolated from the spent cell culture medium by protein G affinity chromatography. The extracellular region of the chCD83 protein was purified and used to immunize mice. A cell fusion was performed, from which 342 hybridomas were screened for mAbs to chCD83. Two mAbs, chCD83-159 and chCD83-227, stained the greatest percentage of chCD83-transfected CHO cells and were selected for further characterization. By flow cytometry, both mAbs reacted with a chicken macrophage cell line, HD11. Both mAbs also recognized a single 53 kDa protein on Western blots of lysates from lipopolysaccharide-stimulated spleen mononuclear cells or unstimulated HD11 cells. Immunostaining of chicken secondary lymphoid organs identified chCD83(+) cells with morphologic and subtissue localization properties comparable to mammalian DCs. In vitro stimulation of spleen mononuclear cells with concanavalin A (Con A) decreased the percentage of chCD83(+) cells compared with cells treated with medium alone. Interestingly, spleen cells treated with Con A in the presence of chCD83-227 mAb exhibited decreased percentage of MHCII(+) cells compared with cells treated with an isotype-matched negative control mAb. These chCD83 mAbs may be useful for future investigations of chicken immune cell maturation and mechanisms of action.     

Comparative analysis of physicochemicals and antioxidative properties in new red rice (Oryza sativa L. cv. Gunganghongmi)

The main objectives of this study were to investigate physicochemicals and antioxidant activities of new red rice (Oryza sativa cv. Gunganghongmi (GH)) by comparing normal brown (Nampyeongbyeo, NB) and reported red rice (Jukjinjubyeo, JB) in Korea. The nutritional constituents, including protein, oil, sugar, fatty acid, GABA, and γ-oryzanol were not significantly different between normal brown and colored rice. However, the ethanol extract of GH showed the highest phenolic content (24.7 ± 1.3 mg g^?1). The ethanol extracts of GH showed higher scavenging activities against DPPH (0.2 mg mL^?1 = 62.1 ± 2.5%) and ABTS (0.2 mg mL^?1 = 63.2 ± 3.5%) radicals. Moreover, GH more inhibited LPS-induced nitric oxide (NO) production (13.2 ± 1.4 μM) than JB (18.3 ± 2.3 μM) and NB (22.1 ± 1.4 μM) at the same concentration (0.2 mg mL^?1) without cytotoxicity. These results suggest that new red rice (GH) would be considered to be new functional rice due to its anti-oxidative effect and high nutrition.     

An unusual case of concomitant infection with chicken astrovirus and group A avian rotavirus in broilers with a history of severe clinical signs

A molecular study of intestinal samples from 21 broiler flocks with a history of enteritis revealed that 23.8% and 14.3% were positive for chicken astrovirus (CAstV) and avian rotavirus (ARV), respectively. CAstV and group A ARV were simultaneously detected in only one broiler flock. Birds in this group developed the significant intestinal lesions characterized by frothy contents, paleness, and thin intestinal walls. In this report we present an unusual case of runting stunting syndrome (RSS) with a history of high mortality and growth retardation in broiler chickens. We also make the first identification of CAstV and group A ARV in broiler chickens in Korea.     

Nasal immunization with major epitope-containing ApxIIA toxin fragment induces protective immunity against challenge infection with Actinobacillus pleuropneumoniae in a murine model

Actinobacillus pleuropneumoniae is an infective agent that leads to porcine pleuropneumonia, a disease that causes severe economic losses in the swine industry. Based on the fact that the respiratory tract is the primary site for bacterial infection, it has been suggested that bacterial exclusion in the respiratory tract through mucosal immune induction is the most effective disease prevention strategy. ApxIIA is a vaccine candidate against A. pleuropneumoniae infection, and fragment #5 (aa. 439–801) of ApxIIA contains the major epitopes for effective vaccination. In this study, we used mice to verify the efficacy of intranasal immunization with fragment #5 in the induction of protective immunity against nasal challenge with A. pleuropneumoniae and compared its efficacy with that of subcutaneous immunization. Intranasal immunization of the fragment induced significantly higher systemic and mucosal immune responses measured at the levels of antigen-specific antibodies, cytokine-secreting cells after antigen exposure, and antigen-specific lymphocyte proliferation. Intranasal immunization not only efficiently inhibited the bacterial colonization in respiratory organs, but also prevented alveolar tissue damage in infectious condition similar to that of a contaminated pig. Moreover, intranasal immunization with fragment #5 provided acquired protective immunity against intranasal challenge with A. pleuropneumoniae serotype 2. In addition, it conferred cross-protection against serotype 5, a heterologous pathogen that causes severe disease by ApxI and ApxII secretion. Collectively, intranasal immunization with fragment #5 of ApxIIA can be considered an efficient protective immunization procedure against A. pleuropneumoniae infection.     

Efficacy of a piglet-specific commercial inactivated vaccine against Porcine circovirus type 2 in clinical field trials

The efficacy of a piglet-specific inactivated Porcine circovirus type 2 (PCV2) vaccine was evaluated with clinical field trials, as recommended by the Republic of Korea’s Animal, Plant & Fisheries Quarantine & Inspection Agency. Three farms were selected on the basis of their history of postweaning multisystemic wasting syndrome. On each farm 60, 1-week-old pigs were randomly allocated to 1 of 2 treatment groups: vaccination at 1 and 3 wk of age or no vaccination. The 2-dose schedule of vaccination with inactivated PCV2 vaccine improved the average daily weight gain from birth to 16 wk of age, the PCV2 load in the blood, and the frequency and severity of lymph node lesions. Inactivated PCV2 vaccine seems to be very effective in controlling PCV2 infection under field conditions.     

Penicillinolide A: A New Anti-Inflammatory Metabolite from the Marine Fungus Penicillium sp. SF-5292

In the course of studies on bioactive metabolites from marine fungi, a new 10-membered lactone, named penicillinolide A (1) was isolated from the organic extract of Penicillium sp. SF-5292 as a potential anti-inflammatory compound. The structure of penicillinolide A (1) was mainly determined by analysis of NMR and MS data and Mosher’s method. Penicillinolide A (1) inhibited the production of NO and PGE₂ due to inhibition of the expression of iNOS and COX-2. Penicillinolide A (1) also reduced TNF-α, IL-1β and IL-6 production, and these anti-inflammatory effects were shown to be correlated with the suppression of the phosphorylation and degradation of IκB-α, NF-κB nuclear translocation, and NF-κB DNA binding activity. In addition, using inhibitor tin protoporphyrin (SnPP), a competitive inhibitor of HO activity, it was verified that the inhibitory effects of compound 1 on the production of pro-inflammatory mediators and NF-κB DNA binding activity were partially associated with HO-1 expression through Nrf2 nuclear translocation.     

Estimating the level of disease risk and biosecurity on commercial poultry farms in New Zealand

ABSTRACT

Aims: To collect baseline data on the contact risk pathways and biosecurity practices of commercial poultry farms in New Zealand, investigate the relationship between the farm-level disease contact risks and biosecurity practices, and identify important poultry health concerns of producers.

Methods: A cross-sectional survey of all registered New Zealand commercial poultry operations was conducted in 2016 collecting information on farm demographics, biosecurity practices, and contact risk pathways. Survey responses were used to generate an unweighted subjective disease risk score based on eight risk criteria and a subjective biosecurity score based on the frequency with which producers reported implementing seven biosecurity measures. Producer opinions towards poultry health issues were also determined.

Results: Responses to the survey response were obtained from 120/414 (29.0%) producers, including 57/157 (36.3%) broiler, 33/169 (19.5%) layer, 24/55 (44%) breeder, and 6/32 (19%) other poultry production types. Median disease risk scores differed between production types (p?<?0.001) and were lowest for breeder enterprises. The greatest risk for layer and broiler enterprises was from the potential movement of employees between sheds, and for breeder enterprises was the on- and off-farm movement of goods and services. Median biosecurity scores also differed between production types (p?<?0.001), and were highest for breeder and broiler enterprises. Across all sectors there was no statistical correlation between biosecurity scores and disease risk scores. Producers showed a high level of concern over effectively managing biosecurity measures.

Conclusions: The uptake of biosecurity measures in the commercial poultry farms surveyed was highly variable, with some having very low scores despite significant potential disease contact risks. This may be related to the low prevalence or absence of many important infectious poultry diseases in New Zealand leading farmers to believe there is a limited need to maintain good biosecurity as well as farmer uncertainty around the efficacy of different biosecurity measures. Further research is needed to understand barriers towards biosecurity adoption including evaluating the cost-effectiveness of biosecurity interventions.