Tiptoeing between restoration and invasion: seed rain into natural gaps within a highly invaded relic forest in the Azores

The last remains of native laurel forest in the Azores are highly threatened by the spread of invasive plants. Because landslides are very frequent in these islands, conservation of native laurel forest requires knowledge of the patterns of bird-dispersed seed rain into forest gaps. We monitored 78 seed traps over 1 year to investigate (1) the role of perches in attracting avian dispersers into gaps, (2) temporal patterns in the dispersal of exotic and native seeds, (3) how seed rain affects vegetation establishment in gaps at different distances from the native forest and (4) whether the caloric content of fruits could explain the number of seeds dispersed. Perches were highly effective in concentrating avian seed dispersal. While some native fruits are produced all year-round, most exotic plants set fruits during the main peak of the native fruit production (August–November). Most seeds recovered from the traps were native, and native seed rain inside the native forest was higher than in gaps. However, deposition of exotic seeds was not affected by distance from native forest. Seed dispersal frequencies monitored by seed traps and by faecal analysis were correlated with each other, but not with fruit caloric content, suggesting that other factors are more important that the nutritional value in predicting avian fruit choice. Forest restoration activities should take into consideration that seed dispersal decreases sharply beyond 100 m from native forest and the attractive potential of perches to direct natural seed dispersal into forest gaps.     

Vaccination with recombinant NetB toxin partially protects broiler chickens from necrotic enteritis

NetB toxin from Clostridium perfringens is a major virulence factor in necrotic enteritis in poultry. In this study the efficacy of NetB as a vaccine antigen to protect chickens from necrotic enteritis was examined. Broiler chickens were immunized subcutaneously with purified recombinant NetB (rNetB), formalin treated bacterin and cell free toxoid with or without rNetB supplementation. Intestinal lesion scores and NetB antibody levels were measured to determine protection after mild oral gavage, moderate in-feed and heavy in-feed challenges with virulent C. perfringens isolates. Birds immunized with rNetB were significantly protected against necrotic enteritis when challenged with a mild oral dose of virulent bacteria, but were not protected when a more robust challenge was used. Bacterin and cell free toxoid without rNetB supplementation did not protect birds from moderate and severe in-feed challenge. Only birds immunized with bacterin and cell free toxoid supplemented with rNetB showed significant protection against moderate and severe in-feed challenge, with the later giving the greatest protection. Higher NetB antibody titres were observed in birds immunized with rNetB compared to those vaccinated with bacterin or toxoid, suggesting that the in vitro levels of NetB produced by virulent C. perfringens isolates are too low to induce the development of a strong immune response. These results suggest that vaccination with NetB alone may not be sufficient to protect birds from necrotic enteritis in the field, but that in combination with other cellular or cell-free antigens it can significantly protect chickens from disease.     

Maternal immunization with vaccines containing recombinant NetB toxin partially protects progeny chickens from necrotic enteritis

Avian necrotic enteritis is a major economic and welfare issue throughout the global poultry industry and is caused by isolates of Clostridium perfringens that produce NetB toxin. Previously we have shown that birds directly vaccinated with inactivated C. perfringens type A culture supernatant (toxoid) combined with recombinant NetB (rNetB) protein were significantly protected from homologous and heterologous challenge. In the present study the protective effect of maternal immunization was examined. Broiler breeder hens were injected subcutaneously with genetically toxoided rNetB(S254L) alone, C. perfringens type A toxoid and toxoid combined with rNetB(S254L). Vaccination resulted in a strong serum immunoglobulin Y response to NetB in hens immunized with rNetB(S254L) formulations. Anti-NetB antibodies were transferred to the eggs and on into the hatched progeny. Subclinical necrotic enteritis was induced experimentally in the progeny and the occurrence of specific necrotic enteritis lesions evaluated. Birds derived from hens immunized with rNetB(S254L) combined with toxoid and challenged with a homologous strain (EHE-NE18) at either 14 or 21 days post-hatch had significantly lower levels of disease compared to birds from adjuvant only vaccinated hens. In addition, birds from hens immunized with rNetB(S254L) alone were significantly protected when challenged at 14 days post-hatch. These results demonstrate that maternal immunization with a NetB-enhanced toxoid vaccine is a promising method for the control of necrotic enteritis in young broiler chickens.     

Evaluation of antiviral activity of phenolic compounds and derivatives against rabies virus

Human rabies is a viral disease with a great impact on public health, mainly on account of its fatal course in the majority of cases. Despite the well-established prophylaxis by immunization, rabies is believed to be responsible for 40,000-70,000 human deaths per year, mostly in endemic areas. Palliative support and experimental protocols to avoid death have been employed with no expressive results, with the exception of a recent human case of recovery from rabies. No antiviral drugs are currently available to fight against this infection. In combination with the prophylaxis, an antiviral drug would be useful for human rabies treatment, providing enhanced protection against the encephalitis caused by the virus. Phenolic compounds are derived from the secondary plant metabolism, although they can also be obtained by synthetic processes. Many studies have shown a great range of pharmacological effects for these substances, including vasodilatation, antiallergenic, antiinflammatory and antiviral properties, among others. In this study, the potential in-vitro anti-rabies activity of 24 synthetic phenolic compounds was evaluated using McCoy cells and PV rabies strain. The cytotoxicity (CC50) was assayed by the MTT method and the antiviral activity (IC50) was estimated by the inhibition of viral cytopathic effects. Isoprinosine and ketamine were used as positive controls. The tested compounds showed selectivity indices (SI=CC50/IC50) ranging from 1.0 to 3.9. Six phenolic compounds failed to inhibit the cytopathic effect to any degree, and four showed SI > or = 3.0. According to these results, some probable structure-activity relationships are suggested. It was observed that the presence of free hydroxyl and ether groups influenced the anti-rabies activity. However, additional studies are required to establish these relationships.     

Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar–gel immunodiffusion (AGID) test for canine brucellosis, were used as the control panel for B. canis infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies.     

The effect of photoperiod and melatonin on plasma prolactin concentrations in female guanaco (Lama guanicoe) in captivity

The present study examined the effects of different photoperiods and melatonin treatment on plasma prolactin concentrations in guanacos, a South American camelid, in captivity. Fourteen adult female guanacos, not gestating or lactating and isolated from males, were studied. The control group was exposed to natural daylight, during short days (N = 7, 10L:14D) and long days (N = 7, 16L:8D). The treatment group (N = 7, 10L:14D) received melatonin implants every 23 days for 6 weeks during long days. Blood samples were taken at intervals of 1 week for 3 weeks, starting the third week of treatment. Prolactin concentrations were measured using competitive ELISA. Plasma concentrations of prolactin in non-lactating female guanacos have seasonal changes, with a higher concentration (p < .001) in short days (3.50 ± 2.24 ng/ml) than long days (1.10 ± 0.91 ng/ml). Melatonin treatment significantly decreases (p < .05) plasma concentrations of prolactin on the 21st day after the treatment. These findings are the first report of an endogenous circannual rhythm of plasma prolactin concentration and the action of melatonin treatment on prolactin secretion in this wild camelid.     

Ruminal and morphometric parameters of the rumen and intestines of sheep fed with increasing levels of spineless cactus (Nopalea cochenillifera Salm-Dyck)

Tropical Animal Health and Production - The objective of this study was to evaluate the ruminal parameters (pH, N-NH3, and microbial protein) and morphometry of the rumen and intestine of sheep fed...     

Co-inoculation of soybeans and common beans with rhizobia and azospirilla: strategies to improve sustainability

Plant–microorganism associations have long been studied, but their exploitation in agriculture partially or fully replacing chemical fertilizers is still modest. In this study, we evaluated the combined action of rhizobial and plant growth-promoting rhizobacteria inoculants on the yields of soybean and common bean. Seed inoculation with rhizobia (1.2?×?10⁶ cells seed^?1) was compared to co-inoculation with Azospirillum brasilense in-furrow (different doses) or on seeds (1.2?×?10⁵ cells seed^?1) in nine field experiments. The best in-furrow inoculant dose was 2.5?×?10⁵ cells of A. brasilense seed^?1 for both crops. Inoculation with Bradyrhizobium japonicum increased soybean yield by an average 222 kg?ha^?1 (8.4 %), and co-inoculation with A. brasilense in-furrow by an average 427 kg?ha^?1 (16.1 %); inoculation always improved nodulation. Seed co-inoculation with both microorganisms resulted in a mean yield increase of 420 kg?ha^?1 (14.1 %) in soybean relative to the non-inoculated control. For common bean, seed inoculation with Rhizobium tropici increased yield by 98 kg?ha^?1 (8.3 %), while co-inoculation with A. brasilense in-furrow resulted in the impressive increase of 285 kg?ha^?1 (19.6 %). The cheaper, more sustainable inoculated treatment produced yields equivalent to the more expensive non-inoculated + N-fertilizer treatment. The results confirm the feasibility of using rhizobia and azospirilla as inoculants in a broad range of agricultural systems, replacing expensive and environmentally unfriendly N-fertilizers.