Effect of summer soil moisture and temperature on the vertical distribution of <Emphasis Type="Italic">Tuber magnatum</Emphasis> mycelium in soil

Tuber magnatum, the Italian white truffle, is the world’s most valuable truffle. Despite the economic importance, its biology and ecology are largely unexplored. This gap of knowledge makes difficult to find reliable methods for its cultivation and to protect and increase the production of the natural productive areas. In this study, the vertical distribution of T. magnatum mycelium in productive soil patches was evaluated using a quantitative PCR (qPCR) approach. Data were then used to develop for the first time a simulation model to predict the mycelial dynamics of T. magnatum at varying soil temperatures and moisture. T. magnatum mycelium was abundant up to 30-cm depth, while the model determined the optimal temperature (20 °C) and water potential (~?0 kPa) for growth of T. magnatum mycelium in soil. Such information could be useful to establish proper irrigation scheduling and to enhance the management of T. magnatum sites, for increasing mycelial growth and fruiting body production.     

Local diversity of native arbuscular mycorrhizal symbionts differentially affects growth and nutrition of three crop plant species

Intact whole native AMF communities occurring across a 100-m-long field were used for the evaluation of plant performance, as determined by the actual fungal species colonizing host roots. The soil from distinct plots within a “hot spot” field was collected to set up 54 experimental units where three different plant species were grown, in order to test whether the whole native AMF communities were able to differentially affect plant growth, to assess the genetic identity of the AMF actually colonizing the tested plants and to analyse their community composition in the different hosts. Molecular analyses revealed that plant growth and nutrition of the crop plants were differentially affected by the diverse native arbuscular mycorrhizal communities colonizing the roots of the three plants, whose performance varied depending on the identity of plant hosts and fungal symbionts, more than on a rich and diversified AMF community. Such results, improving our understanding of AMF distribution at the local scale, represent a starting point allowing the selection, isolation and characterization of the most efficient AMF assemblages to be used as inoculants in sustainable food production systems.     

Influence of Pygoscelis Penguin Colonies on Cu and Pb Concentrations in Soils on the Ardley Peninsula,Maritime Antarctica

Penguins can bioaccumulate metals, a portion of which can be deposited in the environment through organic remains such as excrement, carcasses, and eggshells. In order to determine Cu and Pb concentrations and their relationship to soil, organic matter and grain size were determined in 27 samples collected in zones without penguins, penguin transit zones, and Adelie (Pygoscelis adeliae), Chinstrap (P. antarctica), and Gentoo penguin (P. papua) colonies on the Ardley Peninsula, Maritime Antarctica. An atomic absorption spectrophotometry analysis was carried out, organic matter was determined by loss on ignition, and grain size was measured with a laser diffraction particle size analyzer. The principal component analysis shows a relationship between the variables Cu, Pb, and grain size and areas with penguin presence. Cu concentrations in soils varied among areas (χ², 15.707; p =?0.0004), with higher concentrations in transit zones and penguin colonies (142.63 and 140.79 mg/kg, respectively) than in zones without penguins (83.33 mg/kg). Pb concentrations in soils also varied among areas (χ², 6.5029; p =?0.0387), and were higher in transit zones (5.92 mg/kg) than in the penguin colonies (4.45 mg/kg). Grain size differed significantly among areas (χ², 13.506; p =?0.0012), with higher values in transit zones (avg. 37.38 μm) than in penguin colonies (avg. 26.93 μm) and zones without penguins (avg. 20.72 μm). Organic matter did not differ significantly among the studied zones (χ², 2.0882; p =?0.3520). There is a positive correlation between Cu-Pb (Rho, 0.5532; p =?0.0028), Cu-grain size (Rho, 0.4756; p =?0.0130) and Pb-grain size (Rho, 0.4879; p =?0.0098). The presence of penguins increases Cu concentrations in Antarctic soils due to its bioaccumulation and elimination through excrement; however, the presence of penguins has a minor influence on Pb concentration in soil, probably because this metal is stored efficiently in bones, feathers, and eggshells.     

Evidence of Anaplasma infections in European roe deer (Capreolus capreolus) from southern Spain

Anaplasma spp. (Rickettsiales: Anaplasmataceae) are tick-borne pathogens of veterinary and human importance. The wildlife hosts for these pathogens are not well characterized and may play an important role in the epidemiology of the disease. The objective of this research was to study the infection with A. marginale, A. ovis and A. phagocytophilum in free-ranging European roe deer (Capreolus capreolus) from Cádiz, Andalucía, Spain. Of 17 roe deer tested, 14 (82%) and 5 (29%) had antibodies reactive to Anaplasma spp. and A. phagocytophilum by competitive ELISA and indirect immunofluorescent antibody testing, respectively. Polymerase chain reaction and sequence analysis of Anaplasma major surface protein 4 (msp4) gene was conducted on blood samples from all roe deer examined. Nine (53%) animals had evidence of infection with A. ovis and 3 (18%) were positive for A. phagocytophilum. Concurrent infections were not detected. Despite the presence of A. marginale infections in cattle in the study site (36% msp4 PCR-positive animals), none of the msp4 amplicons from roe deer corresponded to A. marginale sequences. A. ovis msp4 sequences were identical to a genotype previously identified in sheep in Sicily, Italy. Two different A. phagocytophilum genotypes were identified in infected roe deer. This is the first report of roe deer naturally infected with A. ovis. These results demonstrate that roe deer are infected with A. ovis and A. phagocytophilum in Spain and suggest that this species may be involved in the natural cycle of these pathogens in this region, thus acting as potential reservoir for transmission to domestic and wild animals.     

Tumor slices as a model to evaluate doxorubicin in vitro treatment and expression of trios of genes PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2 in canine mammary gland cancer

Background

In women with breast cancer submitted to neoadjuvant chemotherapy based in doxorubicin, tumor expression of groups of three genes (PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2) have classified them as responsive or resistant. We have investigated whether expression of these trios of genes could predict mammary carcinoma response in dogs and whether tumor slices, which maintain epithelial-mesenchymal interactions, could be used to evaluate drug response in vitro.

Methods

Tumors from 38 dogs were sliced and cultured with or without doxorubicin 1 μM for 24 h. Tumor cells were counted by two observers to establish a percentage variation in cell number, between slices. Based on these results, a reduction in cell number between treated and control samples ≥ 21.7%, arbitrarily classified samples, as drug responsive. Tumor expression of PRSS11, MTSS1, CLPTM1 and SMYD2, was evaluated by real time PCR. Relative expression results were then transformed to their natural logarithm values, which were spatially disposed according to the expression of trios of genes, comprising PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2. Fisher linear discrimination test was used to generate a separation plane between responsive and non-responsive tumors.

Results

Culture of tumor slices for 24 h was feasible. Nine samples were considered responsive and 29 non-responsive to doxorubicin, considering the pre-established cut-off value of cell number reduction ≥ 21.7%, between doxorubicin treated and control samples. Relative gene expression was evaluated and tumor samples were then spatially distributed according to the expression of the trios of genes: PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2. A separation plane was generated. However, no clear separation between responsive and non-responsive samples could be observed.

Conclusion

Three-dimensional distribution of samples according to the expression of the trios of genes PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2 could not predict doxorubicin in vitro responsiveness. Short term culture of mammary gland cancer slices may be an interesting model to evaluate chemotherapy activity.     

Clinical and hematological signs associated with dogs naturally infected by Hepatozoon sp. and with other hematozoa: a retrospective study in Uberlândia, Minas Gerais, Brazil

An evaluation was made of the clinical and hematological aspects of 115 dogs infected naturally by Hepatozoon sp. and treated at the Hospital School of the Faculty of Veterinary Medicine in Uberlandia, MG, Brazil. Of the 115 dogs for whom peripheral blood films were evaluated, 89 (77.39%) presented parasitemia by Hepatozoon sp. solely, while 26 (22.61%) had combination of Hepatozoon sp., Babesia sp. and Ehrlichia sp. Young male dogs less than a year old, of undefined breed (UB), were the most commonly affected. Thirty-nine (33.92%) of the dogs were asymptomatic while 76 (66.08%) presented varied clinical symptoms, the most frequent being anorexia, pulmonary alterations, hyperthermia, pale mucosae, apathy and/or prostration, and diarrhea. The majority of hematological alterations were normochromic-normocytic anemia, leukocytosis, neutrophilia, and nuclear deviation of neutrophils to the left (NDNL). The findings of this study confirm that Hepatozoon sp. causes clinical and hematological alterations of varied intensity, which, albeit not specific to canine hepatozoonosis, reinforce the notion that the discovery of the agent in dogs, even with low parasitemia, should be taken into consideration.     

Prevalence of Coxiella burnetti infection in wild and farmed ungulates

The aim of this study was to evaluate by serology and PCR analyses the prevalence of Coxiella burnetti infection in ungulates in Spain. Sera were collected from red deer (Cervus elaphus; n=116), roe deer (Capreolus capreolus; n=39), fallow deer (Dama dama; n=13) and cattle (n=79). Sera were tested for anti-C. burnetii antibody detection by means of an immunofluorescence antibody assay (IFA) and C. burnetii DNA was amplified by PCR in samples from ungulates that had antibodies to phase II antigens. Twenty-nine, 15 and 39 percent of the red deer, roe deer and cattle had antibodies against C. burnetii, respectively. None of the fallow deer sera tested positive. Seroprevalence was statistically higher in farmed than in wild red deer and higher in northern than in southern populations, whereas an inverse pattern was observed for the roe deer. Most of the seropositive animals had only anti-C. burnetii phase II antibodies, thus showing the acute nature of infections in the sampled ungulates. These results show that C. burnetii circulates in wild ungulates in Spain and suggest that they can act as pathogen reservoirs for both domestic animals and humans.     

The effects of prednisone on haemostasis in leishmaniotic dogs treated with meglumine antimoniate and allopurinol

Thirty dogs naturally infected with Leishmania infantum were studied in order to determine the effects of treatment on haemostatic function. The animals were divided randomly into two treatment groups: Group 1 received meglumine antimoniate and allopurinol; Group 2 dogs were given the same treatment plus prednisone. Ten healthy animals were used as untreated controls. Clinical examination and determination of platelet aggregation, coagulation factors and biochemical parameters were undertaken before treatment and after 15, 30 and 60 days. A significant improvement in platelet aggregation was detected after 60 days in Group 1, but only after 15 days in Group 2. In both treated groups, platelet aggregation was lower than in the control group at the end of the study. The results suggest that prednisone may be a useful tool in the treatment of haemostatic disorders during canine leishmaniosis. The potential benefits and risks due to the use of corticosteroids in the treatment of leishmaniosis are discussed.     

PCR identification of Leishmania in diagnosis and control of canine Leishmaniasis

Leishmaniases are endemic in many countries, mainly in rural areas. In Brazil, Leishmania infection is responsible for many cases of Leishmaniases, including recent reports in urban regions. Despite their sensitivity, traditional serological and parasitological methods for detecting Leishmaniases have proven inadequate for species discrimination. This study aimed to identify Leishmania species in biological samples by a fast methodology, avoiding "in vitro" cultivation. Knowledge of the Leishmania species is an important tool in regions where both New World visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) are prevalent. As these new foci appear in areas not traditionally endemic for VL, the main problem is to distinguish between true autochthonous infections and infections acquired in other well-known endemic areas. Since, domestic dogs are known to be the main VL and CL reservoir, they are regularly investigated in endemic areas to prevent, principally, severe and often fatal VL in humans. However, several infected dogs present no clinical signs or clinical signs similar to other canine diseases. Here, we evaluated the ability of PCR to diagnose VL and distinguish L. (L.) chagasi from other Leishmania species in domestic dogs. Samples from 114 dogs from 30 cities (Sao Paulo, Brazil) were divided into two groups: 44 symptomatic and 70 asymptomatic. They were assayed by parasitological methods (culture and microscopic examination) and PCR to determine L. (L.) chagasi, L. (V.) braziliensis; and in some cases, Leishmania spp. Parasitological tests and PCR-L. chagasi were concordant in 105 samples (92%). VL was confirmed in 49 dogs, while 56 had negative results. Of the 114 samples, 9 had discordant results, but were further tested by PCR-Leishmania spp. with positive results. VL was also confirmed in 4 dogs having negative parasitological tests and positive PCR-L. chagasi. Consequently, this PCR was positive for 100% (53/49) of dogs with parasites detected in parasitological tests. Also, PCR demonstrated high specificity detecting 61 dogs negative for VL. Leishmania infection was negative in 56 dogs, and 5 with positive culture and PCR-Leishmania spp. had CL since they were positive in PCR-L. braziliensis. This study shows the importance of including PCR in diagnosis of Leishmaniases by differential diagnosis contributing to the surveillance and control of VL programs.     

Pharmacopuncture Versus Acepromazine in Stress Responses of Horses During Road Transport

Acupuncture has been shown to have the beneficial effect of reducing stress responses in animals and humans. Pharmacopuncture is the injection of subclinical doses of drugs into acupoints to give therapeutic results without side effects. This study compared the effects of injecting the usual dose of acepromazine (ACP; 0.1 mg/kg, intramuscularly [I.M.]) with those of pharmacopuncture (1/10 ACP dose at the governing vessel 1 [GV 1] acupoint) on the stress responses of healthy horses undergoing road transport for 2.5 hours. Four different treatments were applied immediately before loading, with 8 animals/treatment: injection of saline or ACP (0.1 mg/kg, I.M.) at the base of the neck; and injection of saline or 1/10 ACP (0.01 mg/kg) at the GV 1 acupoint. The road transport increased heart rate (HR), respiratory rate, body temperature, and serum cortisol of the untreated horses (injected with saline at the base of the neck). Pharmacopuncture at GV 1 reduced the average HR and transport-induced increase in HR at unloading, without changing the other variables. On the other hand, ACP (0.1 mg/kg) produced significant sedation and reduced the transport-induced increase in respiratory rate but without preventing the stress-induced increase of cortisol. Other acupuncture points and drugs should be tested to verify the beneficial effect of this therapy to reduce stress in horses during road transport.