Evaluation of coral bleaching condition <Emphasis Type="Italic">in situ</Emphasis> using an underwater pulse amplitude modulated fluorometer

ABSTRACT:   Photosynthetic activities of hermatypic corals (28 genera, c . 68 species and c. 92 specimen) were measured in November 2000 at Sekisei lagoon, Okinawa, Japan, using an underwater pulse amplitude modulated fluorometer. Maximum quantum yield (Fv/Fm) ranged from 0.61 to 0.70 and had an average value of 0.664 (standard deviation [SD], 0.0289). Following the measurement of Fv/Fm, quantum yield (Fm'-F)/Fm' at eight-leveled actinic-light intensities were obtained from all specimens. Values of electronic transport rates (ETR) were nearly the same under low photosynthetic active radiation (PAR; µmol quanta/m² per s) condition and varied with increasing PAR. The value of ETR at 100 µmol quanta/m² per s ranged from 21.3 to 30.0 and had an average of 27.8 (SD, 2.09). In August 2001, a coral bleaching event was observed in Sekisei Lagoon. During the event, photosynthetic activities of 21 specimens of eight species (seven genera) were measured. By applying two indicators, Fv/Fm and ETR at 100 µmol quanta/m² per s obtained in August 2000, 18 of 21 specimens were suggested under the bleaching process. In case of Porites lutea , seven colonies were visually evaluated for their condition of bleaching and were found to have two 'normal', three 'slightly bleached' and two 'heavily bleached'. From the density of zooxanthellae, colonies evaluated 'normal' or 'slightly bleached' did not show an apparent difference. However, they were clearly identified by Fm/Fv and ETR at 100 µmol quanta/m² per s.     

RESUME:Studies on the Chemical Control of Aonidiella aurantii (Mask.)

Aonidiella aurantii (Mask.) which was accidentally introduced in Sicily in 1968 has since become a dangerous pest requesting special treatments in citrus groves. Experiments have been carried out on Citrus lemon to evaluate the efficacy of several organophosphorous compounds partly in mixture with white oil and this latter applied sole (cf. table 1). The results obtained against young females are set out in table 2; they show excellent results with almost all compounds whereas the same pesticides had a rather poor effect on mature females (cf. table 3).
It is concluded that the replacement of white oils by organophosphorous compounds is not justified providing that the control is directed against larvae and young females. This means that a proper timing is essential to obtain a satisfactory mortality, thus avoiding the use of detrimental broad acting pesticides.     

Ameliorative Effects of Melatonin against Hydrogen Peroxide‐Induced Oxidative Stress on Boar Sperm Characteristics and Subsequent In Vitro Embryo Development

Melatonin, the major secretory product of the pineal gland, scavenges a variety of reactive oxygen and nitrogen species in vivo and in vitro, indicating that melatonin is a potent function as an antioxidant. The objective of this study was to investigate the effect of melatonin in the presence or absence of hydrogen peroxide (H₂O₂) on sperm characteristics (motility, viability, survival rate, membrane integrity, lipid peroxidation (LPO) and mitochondria activity) and also to examine the developmental rates to the blastocysts stage of porcine oocytes fertilized in vitro with semen treated with or without melatonin (100 nm ) in the presence or absence of H₂O₂ (250 μm ). The sperm were treated with melatonin in the presence or absence of H₂O₂ for 3, 6, 9 and 12 h at 37°C and then analysed for the sperm characteristics. The porcine embryos were produced by in vitro maturation and in vitro fertilization (IVM/IVF) using semen treated with or without melatonin (100 nm ) in the presence or absence of H₂O₂ (250 μm ) for 6 h. The semen characteristics, including motility, viability, survival rate, membrane integrity and mitochondria activity, were higher in the groups that were treated with melatonin in comparison to other groups, irrespective of incubation periods. Malondialdehyde levels in control, melatonin and melatonin + H₂O₂ groups were lower than H₂O₂ only group. A positive correlation was shown among motility, viability, survival rate and membrane integrity, but a negative correlation was observed between LPO and the other evaluation methods. The developmental rates to blastocysts of IVM/IVF porcine oocytes fertilized by semen treated with melatonin were significantly increased compared with any other groups, with the cell number of blastocysts shown to have a similar trend to the developmental rates. These results demonstrate that melatonin can improve the semen characteristics during in vitro storage and support the developmental ability of IVM/IVF embryos in pigs.     

Effect of intravenous injection of acetate on the pancreas of sheep

The effect of an intravenous injection of acetate on plasma insulin and glucagon concentration was examined in conscious sheep. Sodium acetate (312 to 5000 mumol kg-1 bodyweight) injection increased plasma insulin concentrations in a dose-dependent manner. Plasma glucagon concentration was not affected by doses up to 1250 mumol kg-1. Doses of 2500 and 5000 mumol kg-1 produced significant increases (P less than 0.05), but not in a dose related manner. The results of this study indicate that the receptive mechanism of the A cell in the sheep pancreas to acetate might be different from that of the B cell.     

Development of new assessment methods for Acropora coral recruitment using coral settlement devices and holes of marine block

We developed new assessment methods for Acropora coral recruitment using coral settlement devices (CSDs) and holes of a marine block. Both devices were deployed in situ before mass spawning of Acropora corals. The CSDs were sampled after 3–4 months of spawning to measure early recruitment, whereas holes of the marine block were observed underwater or MB was sampled to observe holes after 1 year of spawning to measure the recruitment. By combining the results, we would know the recruitment (visually identified underwater) in reefs, including the amount of early recruitment (identified by stereomicroscopic methods), and the environmental condition of the coral reef. These results would help to predict the future of a reef. Nagura Bay had a high early Acropora recruitment; however, due to competition with algae and sedimentation, the recruitment of juvenile corals was extremely low. The reef would possibly be healthy and sustainable at sites where both the early recruitment and recruitment are high. However, if both are low, the reef will be in a critical condition where urgent restoration will be required.     

Evaluation of serological tests for the diagnosis of Neospora caninum infection in dogs: optimization of cut off titers and inhibition studies of cross-reactivity with Toxoplasma gondii

Diagnosis of Neospora caninum infection in dogs is based on serological assays such as the indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assays (ELISA). This study evaluated two serological tests (IFAT and ELISA) for the detection of IgG antibodies to N. caninum in 300 serum samples of dogs through the optimization of cut off titers by using the two-graph receiver-operating characteristic (TG-ROC) curve. In addition, the identification of major cross-reactive antigens with Toxoplasma gondii was investigated by inhibition ELISA and immunoblotting (IB) assays. IFAT and ELISA results showed 74% agreement, with a good negative concordance (P(neg)=0.83), but a poor positive concordance (P(pos)=0.42). The great majority (86%) of sera with positive concordant results (IFAT+/ELISA+) recognized at least two out of three N. caninum immunodominant antigens, particularly the 29-32 and 35-37 kDa bands. Optimization of cut off titers in IFAT and ELISA was performed considering the reactivity to at least two out of three N. caninum immunodominant antigens as infection markers, obtaining a titer of 50 for IFAT and 200 for ELISA. Seropositivity to N. caninum was significantly associated with T. gondii-seropositive samples, particularly in ELISA (55.4%). Inhibition ELISA curves for N. caninum showed a partial heterologous inhibition, indicating some degree of cross-reactivity between N. caninum and T. gondii antigens. Inhibition IB assays showed a moderate heterologous inhibition for N. caninum antigens above 45-50 kDa. These results indicate that ELISA should be used critically when crude tachyzoite antigen preparations are employed, due to possible cross-reactivity with other related parasites as T. gondii. Also, the cut off dilution of 1:50 in IFAT showed to be the most appropriated for N. caninum serology in dogs. Therefore, we suggest that N. caninum immunodominant antigens, specially the 17 and 29-32 kDa proteins, should be selected markers in serological assays for canine neosporosis.     

Characterization of an endopolygalacturonase Gene cppg1 from Phytopathogenic Fungus Chondrostereum purpureum

Chondrostereum purpureum, a phytopathogenic fungus, produces endopolygalacturonase (endoPG) which has been suggested to have a causal role in the silver-leaf symptom of apple trees. In this paper, we detected C. purpureurn-derived endoPG at the infection sites using ELISA with a polyclonal antibody against endoPG I. A gene encoding endoPG I and its homolog were also isolated from the C. purpureum genome. The endoPG I gene was designated as cppg1. The cppg1 gene is the first fungal endoPG gene reported in the Basidiomycetes. Received 31 May 2000/ Accepted in revised form 13 September 2000     

Assessment of Genetic Variability in the Coding Sequence of Melatonin Receptor Gene (MTNR1A) in Tropical Arid Sheep Breeds of India

Seasonal behaviour in sheep, which varies in tropical and temperate environmental conditions, is a matter of study, because it can provide a clue to address the problem of seasonality in sheep. Melatonin receptor is the membrane‐bound G‐coupled receptor, sensing the message of photoperiodic cues thorough melatonin. Restriction fragment length polymorphism (RFLP) studies were carried out to assess the variability of gene at G612A and C606T SNPs in MTNR1A gene, which have been studied to be markers for out‐of‐season breeding. Allelic frequency distribution corresponded to higher frequency of GG and CC genotype, in tropical arid sheep breed in comparison with temperate region sheep breed. PCR amplification of MTNR1A gene of 30 animals was performed and single nucleotide polymorphisms (SNPs) identification was carried out using Lasergene software. Seven SNPs/mutations were identified, but most of them were synonymous, except the one G706A, leading to substitution of valine by isoleucine. Polyphen‐2 analysis of G706A mutation revealed that it is a benign mutation. Two important SNPs C426T and G555A, which were identified in temperate sheep breeds, could not be traced in Magra and Marwari breeds of sheep. Thus, the Magra and Marwari breeds of tropical, arid region demonstrated the presence of both polymorphic SNPs markers G612A and C606T, associated with out‐of‐season breeding. GG and CC genotypes were having a higher prevalence in the studied population.     

Enrofloxacin is able to control Toxoplasma gondii infection in both in vitro and in vivo experimental models

Currently, toxoplasmosis is treated with sulfadiazine and pyrimethamine. However, this treatment presents several adverse side effects; thus, there is a critical need for the development and evaluation of new drugs, which do not present the same problems of the standard therapy. Enrofloxacin is a fluoroquinolone antibiotic known to control infection against several bacteria in veterinary medicine. Recently, this drug has demonstrated protective effects against protozoan parasites such as Neospora caninum. The present study aimed to determine the effect of enrofloxacin in the control of Toxoplasma gondii infection. For this purpose, human foreskin fibroblast (HFF) cells were infected with T. gondii RH strain and treated with sulfadiazine, penicillin/streptomycin, pyrimethamine, or enrofloxacin. Following treatment, we analyzed the infection index, parasite intracellular proliferation and the number of plaques. Additionally, tissue parasitism and histological changes were investigated in the brain of Calomys callosus that were infected with T. gondii (ME49 strain) and treated with either sulfadiazine or enrofloxacin. Enrofloxacin was able to reduce the infection index, intracellular proliferation and the number of plaques in HFF cells infected by T. gondii in comparison with untreated or penicillin/streptomycin-treated ones. Enrofloxacin was more protective against T. gondii in HFF infected cells than sulfadiazine treatment (P<0.001). In addition, pyrimethamine, enrofloxacin or the associations of sulfadiazine plus pyrimethamine, enrofloxacin plus sulfadiazine or enrofloxacin plus pyrimethamine-treatments were able to reduce the plaque numbers in HFF cells infected by T. gondii when compared to medium, penicillin/streptomycin or sulfadiazine alone. In vivo experiments demonstrated that enrofloxacin diminished significantly the tissue parasitism as well as the inflammatory alterations in the brain of C. callosus infected with T. gondii when compared with untreated animals. Based on our findings, it can be concluded that enrofloxacin is a potential alternative drug for the treatment of toxoplasmosis.