Cloning and purification of the first termicin-like peptide from the cockroach Eupolyphaga sinensis

Termicin is an antimicrobial peptide with six cysteines forming three disulfide bridges that was firstly isolated from the salivary glands and hemocytes of the termite Pseudacanthotermes spiniger. In contrast to many broad-spectrum antimicrobial peptides, termicin is most active against filamentous fungi. Although more than one hundred complementary DNAs (cDNAs) encoding termicin-like peptides have been reported to date, all these termicin-like peptides were obtained from Isoptera insects. Methods The cDNA was cloned by combination of cDNA library construction kit and DNA sequencing. The polypeptide was purified by gel filtration and reversed-phase high performance liquid chromatography (RP-HPLC). Its amino acid sequence was determined by Edman degradation and mass spectrometry. Antimicrobial activity was tested against several bacterial and fungal strains. The minimum inhibitory concentration (MIC) was determined by microdilution tests. Results A novel termicin-like peptide with primary structure ACDFQQCWVTCQRQYSINFISARCNGDSCVCTFRT was purified from extracts of the cockroach Eupolyphaga sinensis (Insecta: Blattodea). The cDNA encoding Es-termicin was cloned by cDNA library screening. This cDNA encoded a 60 amino acid precursor which included a 25 amino acid signal peptide. Amino acid sequence deduced from the cDNA matched well with the result of protein Edman degradation. Susceptibility test indicated that Es-termicin showed strong ability to kill fungi with a MIC of 25 g/mL against Candida albicans ATCC 90028. It only showed limited potency to affect the growth of Gram-positive bacteria with a MIC of 200 g/mL against Enterococcus faecalis ATCC 29212. It was inactive against gram-negative bacteria at the highest concentration tested (400 g/mL). Es-termicin showed high sequence similarity with termicins from many species of termites (Insecta: Isoptera). Conclusions This is the first report of a termicin-like peptide isolated from E. sinensis that belongs to the insect order Blattodea. Our results demonstrate the diversity of termicin-like peptides, as well as antimicrobial peptides in insects.

Performance and nutrient utilization in South American juvenile catfish Pseudoplatystoma spp. weighting 89 - 170g, fed at different energy and protein levels / Desempenho e utilização de nutrientes em juvenis de surubim Pseudoplatystoma spp. alimentados com diferentes níveis de energia e proteína

Most studies in nutrition for the South American catfish (surubim) were limited to the initial phase of development. However, it is clear that performance and nutrient utilization can change during the life stages of a fish. Therefore, this study aimed to evaluate the performance and nutrient utilization in juveniles of surubim fed diets varying in protein and energy levels. Two experiments were performed to test different levels of energy and protein in formulated diets. In the first experiment, surubim juveniles (89.2±4.8g) were fed five diets containing different levels of energy (18.0, 18.8, 19.6, 20.5, 21.3 MJ/kg). In the second experiment, juveniles (170.03±3.35g) were fed five diets containing different levels of protein (360, 400, 440, 480 and 520g/kg). The most favorable energy level for weight gain was 20.3 MJ/kg. The increasing energy levels provided a rise in fat and decrease in protein whole-body composition. The protein amount was between 360 to 400g/kg (383g/kg), which was adequate for performance and nutrient assimilation in surubim juveniles.

A maior parte dos estudos a respeito dos aspectos nutricionais do surubim está limitada às primeiras fases de desenvolvimento. Entretanto, é claro que o desempenho e a utilização dos nutrientes podem mudar durante os diferentes estágios de desenvolvimento destes animais. Assim sendo, este estudo teve como objetivo avaliar o desempenho e a utilização de nutrientes em juvenis de surubim alimentados com dietas contendo níveis variáveis de energia e proteína. Dois experimentos foram realizados para testar os diferentes níveis de proteína e energia. No primeiro experimento, juvenis de surubim (89,2±4,8g) foram alimentados com cinco dietas contendo níveis diferentes de energia (18.0, 18.8, 19.6, 20.5, 21.3MJ/kg). No segundo experimento, os juvenis (170,03±3,35g) foram alimentados com dietas contendo cinco níveis de proteína (360, 400, 440, 480 e 520g/kg). O melhor nível de energia para ganho de peso foi 20,3 MJ/kg. O aumento dos níveis de energia levou a um incremento nos níveis de lipídeo e diminuição da proteína corporal. Níveis de proteína entre 360 a 400g/kg foram os mais adequados para o desempenho e utilização dos nutrientes em juvenis grandes de surubim.

Genetic diversity of porcine reproductive and respiratory syndrome virus in Korea

The high genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV) has been an obstacle to developing an effective vaccine for porcine reproductive and respiratory syndrome (PRRS). This study was performed to assess the degree of genetic diversity among PRRSVs from Korean pig farms where wasting and respiratory syndrome was observed from 2005 to 2009. Samples from 786 farms were tested for the presence of PRRSV using reverse transcription PCR protocol. A total of 117 farms were positive for type 1 PRRSV while 198 farms were positive for type 2. Nucleotide sequences encoding the open reading frame (ORF) 5 were analyzed and compared to those of various published PRRSV isolates obtained worldwide. Sequence identity of the ORF 5 in the isolates was 81.6~100% for type 1 viruses and 81.4~100% for type 2 viruses. Phylogenetic analysis of the ORF 5 sequences showed that types 1 and 2 PRRSVs from Korea were mainly classified into three and four clusters, respectively. The analyzed isolates were distributed throughout the clusters independent of the isolation year or geographical origin. In conclusion, our results indicated that the genetic diversity of PRRSVs from Korean pig farms is high and has been increasing over time.

The signal sequence of type II porcine reproductive and respiratory syndrome virus glycoprotein 3 is sufficient for endoplasmic reticulum retention

The glycoprotein 3 (GP3) of type II porcine reproductive and respiratory syndrome virus has the characteristic domains of a membrane protein. However, this protein has been reported to be retained in the endoplasmic reticulum (ER) rather than transported to the plasma membrane of the cell. In this study, we performed confocal laser scanning microscopy analysis of variants of GP3 and foundthat the signal sequence of the GP3 led to confinement of GP3 in the ER, while the functional ortransmembrane domain did not affect its localization. Based on these results, we concludedthat the signal sequence of GP3 contains the ER retention signal, which might play an important role in assembly of viral proteins.

Frequency of group A rotavirus with mixed G and P genotypes in bovines: predominance of G3 genotype and its emergence in combination with G8/G10 types

The present study describes the genotypic distribution of rotaviruses (RVs) in an Indian bovine population with unexpectedly higher proportions of G3 alone or in combination of G8/G10. PCR-genotyping confirmed that 39.4% (13/33) of the prevalent RVs were the G3 type while 60.6% (20/33) were dual G3G10 or G3G8 types. P typing revealed that 93.9% (31/33) of the samples were P[11] while 6.1% (2/33) possessed a dual P[1]P[11] type. Sequence analysis of the VP7 gene from G3 strains viz. B-46, 0970, and BR-133 showed that these strains had sequence identities of 90.5% to 100% with other bovine G3 strains. The highest identity (98.9% to 100%) was observed with RUBV3 bovine G3 strains from eastern India. The G3 strains (B-46, 0970, and BR-133) showed 97.5% to 98.8% sequence homologies with the Indian equine RV strain Erv-80. Phylogenetic analysis demonstrated that G3 strains clustered with bovine RUBV3 and J-63, and equine Erv-80 G3. Overall, these results confirmed that the incidence of infection by RVs with the G3 genotype and mixed genotypes in the bovine population was higher than previously predicted. This finding reinforces the importance of constantly monitoring circulating viral strains with the G3 genotype in future surveillance studies.