Short communication: Effect of genetic type on antioxidant activity of Caciocavallo cheese during ripening.

The aim of this work was to investigate the antioxidant activity of Caciocavallo cheese made from the milk of 2 breeds, Italian Brown and Italian Holstein, and ripened for 1, 30, 60, 90, and 150 d. The antioxidant activity of cheese was measured using the 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), ferric-reducing antioxidant power (FRAP), and thiol assays. Statistical analysis showed a significant effect of the studied factors. Italian Brown cheese had higher antioxidant activity than Italian Holstein cheese, and antioxidant activity increased during ripening of both cheeses types. Moreover, antioxidant activity varied during ripening depending on the rate of formation of soluble peptides. To date, few studies have evaluated the effect of genetic type on antioxidant capacity of the pasta filata cheeses; thus, this study forms the basis of new knowledge that could lead to the production of a pasta filata cheese with specific nutraceutical characteristics.

Effects of genetic type, stage of lactation, and ripening time on Caciocavallo cheese proteolysis.

The objective of this experiment was to evaluate the effects of genetic type, stage of lactation, and ripening time on proteolysis in Caciocavallo cheese. One hundred twenty Caciocavallo cheeses made from the milk of 2 breeds, Italian Brown and Italian Holstein and characterized by different stages of lactation were obtained and ripened for 1, 30, 60, 90, and 150d. Cheese proteolysis was investigated by ripening index (ratio of water-soluble N at pH 4.6 to total protein, %) and by the study of degradation of the protein fractions (αS1-, ß-, and para-κ-casein), which was determined by densitometric analysis of isoelectric focusing results. The statistical analysis showed a significant effect of the studied factors. Ripening index was higher in Italian Brown Caciocavallo cheese and in cheeses made with early lactation milk, whereas casein solubilization was greater in the first 2mo of ripening. Isoelectric focusing analysis of cheese samples during ripening showed extensive hydrolysis of caseins. In particular, the protein fraction that underwent major degradation by proteolytic enzymes was αS1-casein, followed by ß-casein, whereas para-κ-casein was less degraded. Italian Brown cheese showed a lower residual quantity of ß- and para-κ-casein, whereas Italian Holstein cheese showed a lower residual quantity of αS1-casein. In addition, significant interactions of both first and second order were found on both ripening index and degradation of protein fractions. This study demonstrated that the analyzed factors influenced proteolysis of Caciocavallo cheese, which forms the basis of new knowledge that could lead to the production of a pasta filata cheese with specific characteristics.

Molecular Evolution of Infectious Bronchitis Virus and the Emergence of Variant Viruses Circulating in the United States.

Infectious bronchitis virus (IBV) is a highly infectious and transmissible gammacoronavirus that is nearly impossible to control through biosecurity. Coronaviruses are RNA viruses with an enormous capacity for rapid replication and high rates of mutation, leading to a tremendous amount of genetic diversity. Viral evolution occurs when selection working on genetic diversity leads to new mutations being fixed in the population over time. For IBV, the emergence of variant viruses is likely due to a combination of selection acting on existing genetic diversity, as well as on newly created mutations as the virus replicates, or genetic drift. Immunity against IBV creates a strong selection pressure; however, immunity can also reduce the viral load, decreasing replication and the development of new mutations. Examining the balance between immunity reducing infection, replication, and genetic diversity, and immune pressure selecting for new variants, is extremely difficult at best. Nonetheless, vaccination and immunity do play a role in the emergence of new antigenic variants of IBV. To complicate the situation even more, coronaviruses can undergo recombination, and several studies in the literature report recombination between IBV vaccines and field viruses. However, to our knowledge, unlike genetic drift, recombination alone has not been shown to result in a new antigenic and pathogenic IBV type emerging to cause widespread disease in poultry. Vaccines against IBV that result in an immune population can reduce transmission (basic reproductive number R0 less than 1), making vaccines for IBV the best control strategy available. However, IBV control remains extremely challenging because of the high number of antigenic variants causing disease in poultry and a limited number of vaccines that mostly provide only partial protection against infection and replication of those variants. Currently, there is one major variant IBV circulating in all sectors of US commercial poultry production: DMV/1639/11. This virus was initially detected in 2011, but only began causing significant disease in 2014/2015. Since then, it has affected all three sectors of poultry production (layers, breeders, broilers) and continues to predominate in certain regions of the United States. Additionally, a previously classified variant IBV, which is no longer considered a variant virus, GA08, is highly prevalent. This is attributed to heavy GA08-type IBV vaccine usage because disease caused by the GA08-type virus is rare. Interestingly, the major IBV detected in poultry for several decades, ArkDPI, is no longer among the most detected viruses in the United States. This change corresponds to the shift away from ArkDPI vaccine usage in the broiler sector as GA08 vaccine usage has increased and highlights the role IBV vaccines play in influencing viral populations in commercial chickens.

Estudio recapitulativo- Evolución molecular del virus de la bronquitis infecciosa y aparición de virus variantes que circulan en los Estados Unidos. El virus de la bronquitis infecciosa es un gammacoronavirus altamente infeccioso y transmisible que es casi imposible de controlar mediante bioseguridad. Los coronavirus son virus ARN con una enorme capacidad de replicación rápida y altas tasas de mutación, lo que conduce a una gran cantidad de diversidad genética. La evolución viral ocurre cuando la selección que tiene influencia sobre la diversidad genética conduce a la fijación de nuevas mutaciones en la población a lo largo del tiempo. En el caso del virus de la bronquitis infecciosa, la aparición de variantes virales probablemente se deba a una combinación de selección que actúa sobre la diversidad genética existente, así como a mutaciones recién creadas a medida que el virus se replica o desarrolla deriva genética. La inmunidad contra el virus de la bronquitis infecciosa crea una fuerte presión de selección; sin embargo, la inmunidad también puede reducir la carga viral, disminuyendo la replicación y el desarrollo de nuevas mutaciones. La evaluación del equilibrio entre la inmunidad que reduce la infección, la replicación, la diversidad genética y la presión inmune que selecciona nuevas variantes, es extremadamente difícil en el mejor de los casos. No obstante, la vacunación y la inmunidad desempeñan un papel en la aparición de nuevas variantes antigénicas del virus de bronquitis. Para complicar aún más la situación, los coronavirus pueden someterse a recombinación y varios estudios en la literatura describen acerca de la recombinación entre las vacunas de bronquitis infecciosa y los virus de campo. Sin embargo, hasta donde se conoce, a diferencia de la deriva genética, no se ha demostrado que la recombinación por sí sola dé como resultado nuevos tipos antigénicos o patógenos del virus de la bronquitis infecciosa que causen una enfermedad generalizada en la avicultura. Las vacunas contra el virus de la bronquitis infecciosa que dan como resultado poblaciones inmune pueden reducir la transmisión (número reproductivo básico R0 menor que 1), lo que hace que las vacunas contra bronquitis infecciosa sean la mejor estrategia de control disponible. Sin embargo, el control de la bronquitis infecciosa sigue siendo un gran desafío debido a la gran cantidad de variantes antigénicas que causan enfermedades en la avicultura y a una cantidad limitada de vacunas que, en su mayoría, brindan solo una protección parcial contra la infección y la replicación de esas variantes. Actualmente, existe una variante principal del virus de la bronquitis infecciosa que circula en todos los sectores de la producción avícola comercial de los Estados Unidos: la variante DMV/1639/11. Este virus se detectó inicialmente en 2011, pero solo comenzó a causar una enfermedad significativa entre los años 2014/2015. Desde entonces, ha afectado a los tres sectores de la producción avícola (ponedoras, reproductoras, pollos de engorde) y continúa predominando en ciertas regiones de los Estados Unidos. Además, una variante de este virus previamente clasificada, que ya no se considera una variante, el virus GA08, es muy prevalente. Esto se atribuye al uso intensivo de la vacuna contra este tipo GA08 porque la enfermedad causada por el virus de tipo GA08 es poco común. Curiosamente, el principal virus de la bronquitis detectado en la avicultura durante varias décadas, ArkDPI, ya no se encuentra entre los virus más detectados en los Estados Unidos. Este cambio corresponde a la disminución en el uso de la vacuna ArkDPI en el sector de pollos de engorde a medida que el uso de la vacuna GA08 ha aumentado y se destaca el papel que desempeñan las vacunas de bronquitis infecciosa en la influencia de las poblaciones virales en los pollos comerciales.

Status of research on hydrogen sulphide gas in Chinese mines.

In the process of coal mining, the abnormal gushing of hydrogen sulphide in mines poses a major threat to the health of coal miners and the safe production of mines, as these types of accidents have occurred in many coal-producing countries. China is the largest coal producer and consumer in the world and is one of the countries that are substantially affected by hydrogen sulphide in mines. Based on the existing studies, many investigators in China have conducted research on the actual situation in China and obtained some results. This paper summarizes the previous findings and the current status of hydrogen sulphide in Chinese mines, and predicts the direction of future development. In this paper, we discuss the cause, abnormal distribution, abnormal gushing and prevention and control measures for hydrogen sulphide in mines. In addition, this paper also evaluates the impact of the hydrogen sulphide in mines on the environment and health. This paper outlines a systematic research framework regarding hydrogen sulphide in mines and assesses the interrelationship between subtopics within this system framework. Currently, the scientific research on hydrogen sulphide in mines is not sufficient to meet the needs of the affected individuals Therefore, researchers must increase their efforts in this area to jointly address the challenge of analysing hydrogen sulphide in mines. In addition, we hope that this paper will provide some guidance for the study of hydrogen sulphide in mines.

Comparative Transcriptome Analyses of Longissimus thoracis Between Pig Breeds Differing in Muscle Characteristics.

The two breeds, Mashen (MS; a northern China breed) and Large White (LW; a western lean breed) pigs, show important phenotypic differences in growth, adaptability, intramuscular fat (IMF) content, and energy metabolism since early developmental stage. The main aim of this study was the evaluation of longissimus thoracis muscle transcriptome profile of both genetic types to identify genes, pathways responsible for their differentiated phenotype. Longissimus thoracis of MS and LW pigs were sampled at 0 day (early stage), 90 days (middle stage), and 180 days (later stage) after birth. A total of 3,487 differentially expressed genes (DEGs) were identified at the three time points. Sample clustering analysis revealed the slower growth rate of MS than LW pigs. Gene expression pattern analysis revealed that multicellular organism growth genes (GHSR, EZR, FOXS1, DRD2, SH3PXD2B, CSF1, and TSHR) were involved in the fast growth rate of LW pigs at early stage. Furthermore, DEGs (ACACA, ACSF3, OXSM, CBR4, and HSD17B8) functionally related to fatty acid synthesis revealed that IMF accumulation occurred around 90 and up to 180 days. These DEGs provided valuable resource to study the phenotypic difference in longissimus thoracis muscle between MS and LW pigs.

Blood parameters in fattening pigs from two genetic types fed diet with three different protein concentrations.

The study aimed to evaluate possible differences between two genetic groups (GG) of pigs, fed diets varying in dietary CP level, in hematological and biochemical plasma profiles. The study was carried out in an experimental farm and involved 36 barrows (average BW 129 ± 11 kg) from two GG: group A (18 Italian Duroc boars × Italian Large White sows) and group D (18 DanBred Duroc), fed three experimental diets: a conventional diet and two low-protein diets (LP1 and LP2). A digestibility/balances trial was carried out on 12 pigs A and 12 pigs D that were housed individually in metabolic cages during four digestibility/balances periods. The experimental design was a factorial design, with 3 diets × 2 GG × 4 periods. The experiment lasted 56 d. Blood was sampled from jugular vein in the morning before feed distribution from all barrows in pens at the start and the end of the experimental period; a supplementary blood sample was collected from the 24 pigs at the end of the four digestibility periods (six pigs per period). Blood was analyzed for hematological and biochemical parameters and serum protein profile using automated analyzers. The GG D showed lower white blood cells (WBC), lymphocyte, and monocyte counts than A group. The GG affected several plasma metabolite concentrations: triglycerides, creatinine, Cl, Fe, alkaline phosphatase, and tartrate-resistant acid phosphatase activities were higher in D groups, while urea, albumin, Ca, Na, total bilirubin, and albumin as percentage of total protein were lower than A group. On the contrary, the dietary protein level neither affects WBC nor their populations; only a trend was reported for erythrocytes (red blood cell) and platelets. The diet affected only plasma urea and total bilirubin concentrations.

Long-term implications of feed energy source in different genetic types of reproductive rabbit females. II. Immunologic status.

Genetic selection and nutrition management have played a central role in the development of commercial rabbitry industry over the last few decades, being able to affect productive and immunological traits of the animals. However, the implication of different energy sources in animals from diverse genetic lines achieving such evolutionary success remains still unknown. Therefore, in this work, 203 female rabbits housed and bred in the same conditions were used from their first artificial insemination until their fifth weaning. The animals belonged to three different genetic types diverging greatly on breeding goals (H line, hyper-prolific (n=66); LP line, robust (n=67) and R line, selected for growth rate (n=67), and were assigned to two experimental diets, promoting major differences in energy source (cereal starch or animal fat)). The aims of this work were to: (1) characterize and describe blood leucocyte populations of three lines of rabbit does in different physiological stages during their reproductive period: first artificial insemination, first weaning, second parturition and fifth weaning; and (2) study the possible influence of two different experimental diets on the leucocyte populations in peripheral blood. Flow cytometry analyses were performed on blood samples taken from females at each different sampling stade. Lymphocyte populations at both weanings were characterized by significantly lower counts of total, CD5+ and CD8+ lymphocytes (-19.8, -21.7 and -44.6%; P<0.05), and higher counts of monocytes and granulocytes (+49.2 and +26.2%; P<0.05) than in the other stages. Females had higher blood counts of lymphocytes B, CD8+ and CD25+ and lower counts of CD4+ at first than at fifth weaning (+55.6, +85.8, +57.5, -14.5%; P<0.05). G/L ratio was higher at both weanings (P<0.05), and CD4+/CD8+ ratio increased progressively from the 1AI to the 5 W (P<0.001). Regarding the effect of genetic type in blood leucocyte counts, LP animals presented the highest counts for total, B, CD5+ and CD8+ lymphocytes (+16.7, +31.8, +24.5 and +38.7; P<0.05), but R rabbits showed the highest counts for monocytes and granulocytes (+25.3 and +27.6; P<0.05). The type of diet given during the reproductive life did not affect the leucocyte population counts. These results indicate that there are detectable variations in the leucocyte profile depending on the reproductive stage of the animal (parturition, weaning or none of them). Moreover, foundation for reproductive longevity criteria allows animals to be more capable of adapting to the challenges of the reproductive cycle from an immunological viewpoint.

Characterization of genome segments 2, 3 and 6 of epizootic hemorrhagic disease virus strains isolated in Japan in 1985-2013: Identification of their serotypes and geographical genetic types.

We characterized genome segments 2, 3 and 6 (Seg-2, Seg-3 and Seg-6) of 11 Japanese strains of epizootic hemorrhagic disease (EHD) virus (EHDV) isolated in 1985-2013. The Japanese strains were divisible into two groups based on phylogenetic analyses of the nucleotide sequences of Seg-2 and Seg-6. In both of the phylogenetic trees based on Seg-2 and Seg-6, seven of the 11 Japanese strains were grouped together with EHDV-2 and EHDV-7 strains, and the other four Japanese strains were grouped with EHDV-1 strains. The phylogenetic analysis of Seg-2 among EHDV strains identified 10 of the 11 Japanese strains as EHDV-1, EHDV-2 or EHDV-7. The other Japanese strain, ON-4/B/98, isolated from an asymptomatic cow in 1998 was in the same group as the EHDV-2 and EHDV-7 strains in the phylogenetic trees based on Seg-2 and Seg-6, but the results suggested that the strain belongs to another serotype. We thus conducted a serum neutralization test to identify that serotype by using anti-EHDV-2 and anti-EHDV-7 rabbit sera. We observed that the ON-4/B/98 strain was not sufficiently neutralized by any of the antisera, which suggests that the strain could be assigned into a new serotype, tentatively named 'EHDV-10.' Sequences of Seg-3 were also determined, and all of the Japanese strains were grouped together with Australian strains, suggesting that the Japanese strains are a part of EHDV distributed in the Asia-Pacific region. The data obtained herein would be beneficial for the diagnosis and prevention of EHD in Japan and neighboring countries.

Characterisation of Yersinia pseudotuberculosis isolated from animals with yersiniosis during 1996-2013 indicates the presence of pathogenic and Far Eastern strains in Italy.

Yersinia pseudotuberculosis is a pathogen that infects both animals and humans worldwide. The epidemiology of infection caused by Y. pseudotuberculosis is poorly understood; however, its outbreaks have been traced back to a probable source in wildlife. This study aimed to characterise Y. pseudotuberculosis isolates collected from animals with yersiniosis. This study included 90 isolates of Y. pseudotuberculosis collected from different animals with yersiniosis between 1996 and 2013 in Italy. The isolates were tested for antimicrobial susceptibility and were biotyped. Genes associated with virulence plasmid pYV and those encoding O-antigen, high pathogenicity island (HPI), and superantigenic toxin (YPM) were determined by performing PCR. Pulsed-field gel electrophoresis (PFGE) was performed using NotI and SpeI enzymes, and 3 dendrograms were generated. No antibiotic resistance was found. The presence of pYV was shown in 57 out of 90 isolates. Virulence profiles of majority of the isolates indicated that they belonged to O:1a and O:1b serotypes, biotype 1, and genetic type 2. Isolates belonging to O:2a serotype were detected in sheep and cattle and were found to be associated (for the first time) with septicemia in hares. Y. pseudotuberculosis isolates belonging to O:5a and O:12-O13 serotypes were also detected in hares. To our knowledge, this is the first study to detect Y. pseudotuberculosis isolates belonging to the O:12-O13 serotype from a clinical case in Europe. Results of PFGE indicated that it was a reliable method for investigating the genetic relatedness of Y. pseudotuberculosis isolates. Thus, characterisation of Y. pseudotuberculosis infection in animals should be considered a possible tool for the surveillance of pseudotuberculosis.

Determination of Globin Beta-chains and Comparison of Genetic Type of SMMC/B Mice of Three Generations by Peptide Map Analysis / 第二军医大学学报

The separation and purification of beta chains of globm from the inbreeding SMMC/B mice were performed by the dissociation of chains in urea, CMhcellulose column chromatography for the separation of alpha and beta chains, and Sephadex G25 column chromatography for removing salts. The pure beta-chain was digested by trypsin. Digested fragments were resolved by cellulose membrane two-way electrophoresis, thus obtaining the map of spots of beta-chain fragments for B33, B34, Bp(), Bp and Bf. The spots,BTp3, which have the same positions ,were separated and micro-sequences were assayed by DABITC/PITC double coupling technique. According to the genetic code, we might deduce the nucleotide sequence of their mRNA from the known amino acid sequence of BTp3, a piece of beta-chain. By analyzing the altered genetic loci of beta-chain of globin, we might determine the genetic purity of the strain and distinguish objectively the genetic types between species and subspecies.