Ecological evaluation of polymetallic soil quality: the applicability of culture-dependent methods of bacterial communities studying

Purpose

This work aimed to study the effect of long-term polymetallic contamination on the state and parameters of soil bacterial communities, including the abundance of different groups of culturable bacteria and the activity of nitrification.

Materials and methods

Monitoring plots were located in the dry lake and surrounding area, which had been formerly used for the discharge of industrial waste. The soils in the 16 plots were characterized by extremely high levels of heavy metal pollution. This study evaluated the main soil physicochemical properties by various methods, total metal contents by X-ray analysis, mobile metal content by atomic absorption spectrophotometry, the abundance of chosen groups of culturable bacteria by inoculation on solid media, and nitrification activity from ammonium and nitrite oxidation rates.

Results and discussion

High adaptation capacity of microbial communities to long-term pollution was revealed through marked lack of decrease in the abundance of some of the bacterial groups in soils with high contamination levels. Among the bacteria determined by the colony count method, copiotrophic and spore-forming bacteria were the least sensitive to contamination, and actinomycetes were the most sensitive. The high levels of soil pollution with heavy metals had pronounced adverse effects on nitrification activity. The decrease in activity was strongly correlated with pollutant concentrations. The oxidation of nitrite was shown to be more affected by pollution that the oxidation of ammonium.

Conclusions

Some groups and parameters of culturable microorganisms can be used for soil status estimation under pollution conditions even though they are only a small fraction of the microbial community. The most sensitive parameter was the nitrification rate, while the number of actinomycetes was found to be most promising parameter among the groups of bacteria determined by plate counts. The use of sensitive groups of culturable microorganisms for bioindication purposes is a method, which may provide a cheap and sufficiently reliable tool for large-scale soil monitoring studies.

     

A survey of mutations in the Cochliomyia hominivorax (Diptera: Calliphoridae) esterase E3 gene associated with organophosphate resistance and the molecular identification of mutant alleles

Cochliomyia hominivorax (Calliphoridae) is one of the most important myiasis-causing flies and is responsible for severe economic losses to the livestock industry throughout the Neotropical region. In Brazil, C. hominivorax has been controlled mainly with organophosphate (OP) insecticides, although the inappropriate use of these chemicals can result in the selection of resistant flies. Changes in carboxylesterase activity have been associated with OP insecticides in some arthopodan species. In this work, we isolated and characterized part of the E3 gene in C. hominivorax (ChE7), which contained the same substitutions responsible for the acquisition of OP hydrolase activity in Lucilia cuprina (Calliphoridae). Digestion of the polymerase chain reaction products with a restriction enzyme that specifically recognized the mutation site unambiguously differentiated wild and mutated esterase alleles. The PCR-RFLP assay therefore provided a fast, reliable DNA-based method for identifying C. hominivorax individuals with a mutation in the esterase gene. Further bioassays to determine the association of this mutation with OP resistance in C. hominivorax should allow the development of more effective strategies for managing this species.     

Detection of porcine circovirus genotypes 2a and 2b in aborted foetuses from infected swine herds in the State of S?o Paulo,Brazil

Background

Porcine circovirus type 2 (PCV2) has been associated with several disease complexes, including reproductive failure. The aim of this study was to identify the subtypes of PCV2 that are associated with reproductive failure in pigs from the State of São Paulo, Brazil and to investigate co-infections with other infectious organisms.

Findings

Samples of 168 aborted foetuses or mummified foetuses from five farrow-to-finish swine farms known to be infected with PCV2 and located in the State of São Paulo were tested for PCV2 by polymerase chain reaction (PCR). Positive samples were additionally tested for porcine parvovirus (PPV), Leptospira spp. and Brucella spp. by PCR. PCV2 was detected in 18 of the samples (10.7%). PPV, Brucella spp. and Leptospira spp were found in 2, 10 and 0 cases, respectively. Eleven PCV2 strains were sequenced and determined to be either genotype 2a (n = 1) or 2b (n = 10).

Conclusions

The findings indicate that the frequency of PCV2 infections in aborted porcine foetuses from the State of São Paulo is rather low (10.7%) and that co-infection with other pathogens is common and may be involved in PCV2 associated reproductive failure. No repeatable, characteristic amino acid motifs for regions of the PCV2 capsid protein seemed to be associated with abortion in sows.     

Serologic and molecular characterization of tickborne pathogens in lions (Panthera leo) from the Fasano Safari Park, Italy.

Lions (Panthera leo) are an endangered species threatened by illegal hunting, habitat loss, and infectious diseases. Little is known about the tick-borne pathogens that infect lions and could contribute to population declines. The objective of this study was to characterize Rickettsia spp., Anaplasma phagocytophilum, and Coxiella burnetii infections in 10 lions from the Fasano Safari Park in Italy by serology, polymerase chain reaction, and sequence analysis. Although animals did not show clinical signs of tick-borne diseases, evidence of infection with C. burnetii, spotted fever group Rickettsia sp., and A. phagocytophilum were found in 50%, 20%, and 10% of the lions, respectively. One of the lions tested positive for all three pathogens. This study is the first report of molecular evidence of infection with C. burnetii, Rickettsia sp., and A. phagocytophilum in lions and provides evidence that these felids become infected and serve as hosts for tick-transmitted bacteria.     

The importance of manual white blood cell differential counts and platelet estimates in elephant hematology: blood film review is essential

Unique features of elephant hematology are known challenges in analytical methodology like two types of monocytes typical for members of the Order Afrotheria and platelet counts of the comparatively small elephant platelet. To investigate WBC differential and platelet data generated by an impedance-based hematology analyzer without availability of validated species-specific software for recognition of elephant WBCs and platelets, compared to manual blood film review. Blood samples preserved in ethylenediaminetetraacetic acid (EDTA) of 50 elephants (n = 35 Elephas maximus and n = 15 Loxodonta africana) were used. A Mann-Whitney test for independent samples was used to compare parameters between methods and agreement was tested using Bland-Altman bias plots. All hematological variables, including absolute numbers of heterophils, lymphocytes, monocytes, eosinophils, basophils, and platelets, were significantly different (p < 0.0001) between both methods of analysis, and there was no agreement using Bland-Altman bias plots. Manual review consistently produced higher heterophil and monocyte counts as well as platelet estimates, while the automated analyzer produced higher lymphocyte, eosinophil, and basophil counts. The hematology analyzer did not properly differentiate elephant lymphocytes and monocytes, and did not accurately count elephant platelets. These findings emphasize the importance of manual blood film review as part of elephant complete blood counts in both clinical and research settings and as a basis for the development of hematological reference intervals.     

Comparison of temperature readings from a percutaneous thermal sensing microchip with temperature readings from a digital rectal thermometer in equids

OBJECTIVE: To compare temperature readings from an implantable percutaneous thermal sensing microchip with temperature readings from a digital rectal thermometer, to identify factors that affect microchip readings, and to estimate the sensitivity and specificity of the microchip for fever detection. DESIGN: Prospective study. ANIMALS: 52 Welsh pony foals that were 6 to 10 months old and 30 Quarter Horses that were 2 years old. PROCEDURES: Data were collected in summer, winter, and fall in groups 1 (n = 23 ponies), 2 (29 ponies), and 3 (30 Quarter Horses), respectively. Temperature readings from a digital rectal thermometer and a percutaneous thermal sensing microchip as well as ambient temperature were recorded daily for 17, 23, and 19 days in groups 1 through 3, respectively. Effects of ambient temperature and rectal temperature on thermal sensor readings were estimated. Sensitivity and specificity of the thermal sensor for detection of fever (rectal temperature, >or= 38.9 degrees C [102 degrees F]) were estimated separately for data collection at ambient temperatures 15.6 degrees C. RESULTS: Mean ambient temperatures were 29.0 degrees C (84.2 degrees F), -2.7 degrees C (27.1 degrees F), and 10.4 degrees C (50.8 degrees F) for groups 1 through 3, respectively. Thermal sensor readings varied with ambient temperature and rectal temperature. Rectal temperatures ranged from 36.2 degrees to 41.7 degrees C (97.2 degrees to 107 degrees F), whereas thermal sensor temperature readings ranged from 23.9 degrees (75 degrees F) to 42.2 degrees C (75 degrees to 108 degrees F). Sensitivity for fever detection was 87.4%, 53.3%, and 58.3% in groups 1 to 3, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: The thermal sensor appeared to have potential use for initial screening of body temperature in equids at ambient temperatures > 15.6 degrees C.