Use of peas in organic buffalo farming: effects on nutrient digestibility and milk production

Twenty lactating buffalo cows, organically farmed, were used to examine the effects of including peas in total mixed ration. Two concentrates were formulated to contain, as the main protein sources, either 350 g/kg of soybean cake (CC) or 450 g/kg of peas (ExpC). Cows were blocked into two groups according to parity and previous milk yield and were assigned to one of two dietary treatments: one group was fed a diet with 6 kg/d of CC, whereas the treatment group was fed diet in which 3kg/d of CC were replaced by an equal quantity of ExpC. Digestibility of the diets and milk production of the cows were measured. The experimental period covered the whole lactation period. No differences were observed between groups for milk yield and composition, and for total tract apparent digestibility. The main hypothesis tested, that the replacement of soybean cake with peas in buffalo diet would not affect milk yield and composition, was confirmed. This suggested that the partial substitution of soybean cake with peas in diet for buffalo cows can be possible without affect performances

Feeding management and milk production in organic and conventional buffalo farms

The feeding management, milk yield and milk composition were investigated in two adjacent buffalo farms, one organic certified (on average, 220 lactating buffalo cows) and one conventional (on average, 314 lactating buffalo cows) located in the Sele Plain (southern Italy). Milk samples from the two farm were collected twice a month during the period from June to November 2006. Milk production was also recorded. The investigated milk components were the content of protein, fat, lactose, urea and the number of somatic cells. The following features about the feeding management of lactating buffalo cows were recorded during monthly farm visits: feed used, herbage utilization, ration composition. Fat and protein correct milk yield was higher in conventional farm than in organic one, but milk components were similar between the farms. The greatest differences were found in the somatic cell count, lower in organic milk than in conventional one, and the urea content, which was higher in organic milk than in conventional farm, but still within the normal range reported for buffalo milk

Effect of Saccharomyces cerevisiae Live Cells on In Vivo Digestibility and Nitrogen Excretion in Lactating Buffaloes

The effects of dietary inclusion of Saccharomyces cerevisiae culture on intake, in vivo digestibility, and fecal nitrogen excretion were examined in dairy buffalo. Forty lactating buffalo cows were equally divided into Control and Saccharomyces groups, balanced for milk production, parity, and days in milk. Two subsequent 16-d experimental phases were carried out. For both groups during the first experimental period a TMR based on maize silage (maize-TMR) was used, whereas in the second period an alfalfa haylage (alfalfa-TMR) was administered to the animals. In each experimental period, Saccharomyces group was supplemented with 50 g/head/day of yeast (Biocell®, Limena, Padova, Italy), corresponding to 20—109 CFU/head/day Saccharomyces cerevisiae NCYC Sc47 strain. The yeast supplement was top-dressed onto the morning feed. Dry matter intake (DMI) was assessed for 6 consecutive d on group basis, by the difference between feed offered and refused. In the last 3 days of experimental period in vivo digestibility was determined by using acid-insoluble ash (AIA) as an intrinsic digestibility marker. Saccharomyces supplemented buffalo cows presented greater DMI of maize-TMR, whereas no statistical differences between the groups were observed for alfalfa-TMR. Saccharomyces supplementation significantly improved in vivo digestibility of both TMR. Fecal nitrogen excretion was significantly reduced by the use of yeast supplementation. Results suggest that the inclusion of Saccharomyces cerevisiae culture in the diet for lactating buffaloes can be recommended for its effects on cow's digestive efficiency and fecal nitrogen excretion

Moving Buffalo Farming beyond Traditional Areas: Performances of Animals, and Quality of Mozzarella and Forages

An observational case study was designed to highlight issues associated with a possible expansion of dairy buffalo (Bubalus bubalis) farming outside the traditional coastal plains of southern Italy. Twenty pregnant buffaloes were transferred to a hilly inland farm. After calving, production and reproduction data were collected monthly throughout lactation. From 4 to 6 months of lactation, buffaloes were enrolled in a feeding trial to evaluate the effects of locally grown forages (maize silage vs. hay) on milk production and in vivo digestibility. Sensory properties of mozzarella cheese produced at a local dairy were also evaluated. No obvious effects of diet were found. Compared to the data recorded in the previous lactation completed in the farm of origin, milk yield was reduced by 37.2%, and milk protein by 6.1%, whereas milk fat improved (+4.5%). A lower pregnancy rate (−13.3%), increased days open (+122%), and a prolonged intercalving period (+26.9%) were also observed. Lactation length was shorter than the standard value of 270 d. The results showed that peculiar reproductive characteristics, lower environmental temperatures, and the specificity of the mozzarella production process are the main problems to be addressed in an expansion of buffalo farming outside traditional area

The Plasmodium berghei serine protease PbSUB1 plays an important role in male gamete egress.

The Plasmodium subtilisin-like serine protease SUB1 is expressed in hepatic and both asexual and sexual blood parasite stages. SUB1 is required for egress of invasive forms of the parasite from both erythrocytes and hepatocytes, but its subcellular localisation, function, and potential substrates in the sexual stages are unknown. Here, we have characterised the expression profile and subcellular localisation of SUB1 in Plasmodium berghei sexual stages. We show that the protease is selectively expressed in mature male gametocytes and localises to secretory organelles known to be involved in gamete egress, called male osmiophilic bodies. We have investigated PbSUB1 function in the sexual stages by generating P. berghei transgenic lines deficient in PbSUB1 expression or enzyme activity in gametocytes. Our results demonstrate that PbSUB1 plays a role in male gamete egress. We also show for the first time that the PbSUB1 substrate PbSERA3 is expressed in gametocytes and processed by PbSUB1 upon gametocyte activation. Taken together, our results strongly suggest that PbSUB1 is not only a promising drug target for asexual stages but could also be an attractive malaria transmission-blocking target

Serum antibody response to Human papillomavirus (HPV) infections detected by a novel ELISA technique based on denatured recombinant HPV16 L1, L2, E4, E6 and E7 proteins

BACKGROUND: Human papillomaviruses (HPVs) are the primary etiological agents of cervical cancer and are also involved in the development of other tumours (skin, head and neck). Serological survey of the HPV infections is important to better elucidate their natural history and to disclose antigen determinants useful for vaccine development. At present, the analysis of the HPV-specific antibodies has not diagnostic value for the viral infections, and new approaches are needed to correlate the antibody response to the disease outcome. The aim of this study is to develop a novel ELISA, based on five denatured recombinant HPV16 proteins, to be used for detection HPV-specific antibodies. METHODS: The HPV16 L1, L2, E4, E6 and E7 genes were cloned in a prokaryotic expression vector and expressed as histidine-tagged proteins. These proteins, in a denatured form, were used in ELISA as coating antigens. Human sera were collected from women with abnormal PAP smear enrolled during an ongoing multicenter HPV-PathogenISS study in Italy, assessing the HPV-related pathogenetic mechanisms of progression of cervical cancer precursor lesions. Negative human sera were collected from patients affected by other infectious agents. All the HPV-positive sera were also subjected to an avidity test to assess the binding strength in the antigen-antibody complexes. RESULTS: Most of the sera showed a positive reactivity to the denatured HPV16 proteins: 82% of the sera from HPV16 infected women and 89% of the sera from women infected by other HPV genotypes recognised at least one of the HPV16 proteins. The percentages of samples showing reactivity to L1, L2 and E7 were similar, but only a few serum samples reacted to E6 and E4. Most sera bound the antigens with medium and high avidity index, suggesting specific antigen-antibody reactions. CONCLUSION: This novel ELISA, based on multiple denatured HPV16 antigens, is able to detect antibodies in women infected by HPV16 and it is not genotype-specific, as it detects antibodies also in women infected by other genital HPVs. The assay is easy to perform and has low cost, making it suitable for monitoring the natural history of HPV infections as well as for detecting pre-existing HPV antibodies in women who receive VLP-based HPV vaccination

Egress-related osmiophilic bodies

© 2014 John Wiley & Sons Ltd. Summary: Gametogenesis is the earliest event after uptake of malaria parasites by the mosquito vector, with a decisive impact on colonization of the mosquito midgut. This process is triggered by a drop in temperature and contact with mosquito molecules. In a few minutes, male and female gametocytes escape from the host erythrocyte by rupturing the parasitophorous vacuole and the erythrocyte membranes. Electron-dense, oval-shaped organelles, the osmiophilic bodies (OB), have been implicated in the egress of female gametocytes. By comparative electron microscopy and electron tomography analyses combined with immunolocalization experiments, we here define the morphological features distinctive of male secretory organelles, hereafter named MOB (male osmiophilic bodies). These organelles appear as club-shaped, electron-dense vesicles, smaller than female OB. We found that a drop in temperature triggers MOB clustering, independently of exposure to other stimuli. MDV1/PEG3, a protein associated with OB in Plasmodium berghei females, localizes to both non-clustered and clustered MOB, suggesting that clustering precedes vesicle discharge. A P.berghei mutant lacking the OB-resident female-specific protein Pbg377 displays a dramatic reduction in size of the OB, accompanied by a delay in female gamete egress efficiency, while female gamete fertility is not affected. Immunolocalization experiments indicated that MDV1/PEG3 is still recruited to OB-remnant structures

Generation of human antibody fragments recognizing distinct epitopes of the nucleocapsid (N) SARS-CoV protein using a phage display approach

BACKGROUND: Severe acute respiratory syndrome (SARS)-CoV is a newly emerging virus that causes SARS with high mortality rate in infected people. Successful control of the global SARS epidemic will require rapid and sensitive diagnostic tests to monitor its spread, as well as, the development of vaccines and new antiviral compounds including neutralizing antibodies that effectively prevent or treat this disease. METHODS: The human synthetic single-chain fragment variable (scFv) ETH-2 phage antibody library was used for the isolation of scFvs against the nucleocapsid (N) protein of SARS-CoV using a bio panning-based strategy. The selected scFvs were characterized under genetics-molecular aspects and for SARS-CoV N protein detection in ELISA, western blotting and immunocytochemistry. RESULTS: Human scFv antibodies to N protein of SARS-CoV can be easily isolated by selecting the ETH-2 phage library on immunotubes coated with antigen. These in vitro selected human scFvs specifically recognize in ELISA and western blotting studies distinct epitopes in N protein domains and detect in immunohistochemistry investigations SARS-CoV particles in infected Vero cells. CONCLUSION: The human scFv antibodies isolated and described in this study represent useful reagents for rapid detection of N SARS-CoV protein and SARS virus particles in infected target cells

Integrase defective lentiviral vector as a vaccine platform for delivering Influenza antigens

Viral vectors represent an attractive technology for vaccine delivery. We exploited the integrase defective lentiviral vector (IDLV) as a platform for delivering relevant antigens within the context of the ADITEC collaborative research program. In particular, Influenza virus hemagglutinin (HA) and nucleoprotein (NP) were delivered by IDLVs while H1N1 A/California/7/2009 subunit vaccine (HAp) with or without adjuvant was used to compare the immune response in a murine model of immunization. In order to maximize the antibody response against HA, both IDLVs were also pseudotyped with HA (IDLV-HA/HA and IDLV-NP/HA, respectively). Groups of CB6F1 mice were immunized intramuscularly with a single dose of IDLV-NP/HA, IDLV-HA/HA, HAp alone or with HAp together with the systemic adjuvant MF59. Six months after the vaccine prime all groups were boosted with HAp alone. Cellular and antibody responses to influenza antigens were measured at different time points after the immunizations. Mice immunized with HA-pseudotyped IDLVs showed similar levels of anti-H1N1 IgG over time, evaluated by ELISA, which were comparable to those induced by HAp+MF59 vaccination, but significantly higher than those induced by HAp alone. The boost with HAp alone induced an increase of antibodies in all groups, and the responses were maintained at higher levels up to 18 weeks post-boost. The antibody response was functional and persistent overtime, capable of neutralizing virus infectivity, as evaluated by hemagglutination inhibition and microneutralization assays. Moreover, since neuraminidase (NA)-expressing plasmid was included during IDLV preparation, immunization with IDLV-NP/HA and IDLV-HA/HA also induced functional anti-NA antibodies, evaluated by Enzyme-Linked Lectine Assay (ELLA). IFN?-ELISPOT showed evidence of HA-specific response in IDLV-HA/HA immunized animals and persistent NP-specific CD8+ T cell response in IDLV-NP/HA immunized mice. Taken together our results indicate that IDLV can be harnessed for producing a vaccine able to induce a comprehensive immune response, including functional antibodies directed towards HA and NA proteins present on the vector particles in addition to a functional T cell response directed to the protein transcribed from the vector

Clinical and epidemiological correlates of antibody response to human papillomaviruses (HPVs) as measured by a novel ELISA based on denatured recombinant HPV16 late (L) and early (E) antigens

<p>Abstract</p> <p>Background</p> <p>At present, seroreactivity is not a valuable parameter for diagnosis of Human Papillomavirus (HPV) infection but, it is potentially valuable as marker of viral exposure in elucidating the natural history of this infection. More data are needed to asses the clinical relevance of serological response to HPV.</p> <p>Objectives</p> <p>The objective was to assess the clinical and epidemiological correlates of HPV-seroreactivity in a cohort of HIV-negative and HIV-positive women.</p> <p>Methods</p> <p>Seroreactivity of 96 women, evaluated in an ELISA test based on denatured HPV16 late (L) and early (E) antigens, was correlated with their clinical and epidemiological data previously collected for a multi-centre Italian study, HPV-PathogenISS study.</p> <p>Results</p> <p>No significant correlation was found between HPV DNA detection and seroreactivity. Women, current smokers showed significantly less seroreactivity to L antigens as compared with the non-smokers. HIV-positive women showed significantly less (66.7%) antibody response as compared with HIV-negative women (89.3%), with particularly impaired response to L antigens. Women, HIV-positive and current smokers, showed by far the lowest seroprevalence (33.3%) as compared to 75.9% among all other women (OR = 0.158; 95%CI 0.036–0.695, p = 0.014; Fisher's exact test). Importantly, this association did not loose its significance when controlled for confounding from age (continuous variable) in multivariate analysis or using Mantel-Haenszel test for age-groups.</p> <p>Conclusion</p> <p>It is tempting to speculate that HIV-positive current smokers comprise a special high-risk group, with highly impaired immunological response that could prevent eradication of persistent HPV infections and thus contribute to development of CIN3/CC.</p