Learning interspecific communicative responses in Pampas foxes (Lycalopex gymnocercus)

Domestic dogs show remarkable communicative abilities in their interaction with people. These skills maybe explained by the interaction between the domestication process and learning experiences during ontogeny. Studies carried out on other species of canids, which have not been domesticated are relevant to this topic. The purpose of this article is to study the effect of instrumental learning on captive Pampas foxes’ (Lycalopex gymnocercus) communicative responses to humans. Seven foxes were tested in a conflict situation involving food within sight but out of their reach. In these situations dogs typically gaze at the human face to ask for food. In Study 1, there was an increase in gaze duration as a consequence of reinforcement and a decrease during extinction, when animals did not receive any more food. In Study 2, all of the four foxes tested successfully followed proximal pointing gesture to find hidden food. When a distal pointing cue was given, three out of four followed it in the first session and one in the second session. These results are consistent with those previously found in dogs, and suggest that learning experiences allow the development of communicative skills, even in not domesticated canid species.     

Chickens (Gallus domesticus) are natural intermediate hosts of Neospora caninum

Neospora caninum naturally infects many mammal species, but has not previously been demonstrated in birds. We examined sera for N. caninum antibodies from 200 outdoor chickens and from 200 chickens confined indoors in the state of Bahia, Brazil. Seroprevalence was greater in outdoor chickens (23.5% versus 1.5%, P<0.001). PCR testing for N. caninum was positive in six of 10 seropositive chickens. Amplicons from two of these were sequenced and had 97-98% nucleotide identity with N. caninum. This finding extends the list of intermediate hosts of N. caninum to include birds and may have important epidemiological consequences.     

Activity,habitat use,density, and reproductive biology of the crab-eating fox (Cerdocyon thous) and comparison with the pampas fox (Lycalopex gymnocercus) in a Restinga area in the southern Brazilian Atlantic Forest

The ecology and reproductive biology of the crab-eating fox (Cerdocyon thous) and the pampas fox (Lycalopex gymnocercus) were studied in Itapuã State Park, an Atlantic Forest fragment. In this study we describe their activity patterns (diel, lunar, and seasonal), reproductive period, density, spatial distribution, and habitat overlap. The crab-eating fox was more abundant than the pampas fox, occurring in all types of habitats and overlapping with the pampas fox only in the southern portion of the study area, covered by open vegetation. Both canid species presented nocturnal habits and C. thous showed no significant differences in activity among lunar phases or else year season. The density of C. thous was estimated to be 0.78 ind/km². The crab-eating fox reproduced once a year, generating pups during spring (October to December). In comparison with C. thous populations from lower-latitude regions the relatively low density and one reproductive event per year that we observed are probably related to a colder climate, shorter photoperiod, and shortage of food resources in the winter, characteristic of southern Brazil.     

Monoclonal antibody inhibition of Neospora caninum tachyzoite invasion into host cells

Monoclonal antibodies were produced against Neospora caninum tachyzoites to identify antigens which may play a role during invasion of host cells. Confocal laser microscopy showed that most antigens recognised by the mAb were located on the surface, but one mAb, 1A5, reacted to the apical end of the parasite. Some mAbs, which recognised 70, 42 and 36kDa parasite proteins, significantly inhibited the invasion of the parasite in vitro. The mAbs which recognised 42 and 36kDa parasite protein, reacted with Nc-p43 and Nc-p36 expressed by vaccinia virus and Escherichia coli, respectively. These results suggest that a 70kDa protein, Nc-p43 and Nc-p36 are involved in the invasion of the parasite into host cells.     

Neospora caninum (Apicomplexa) in an aborted equine fetus

Tachyzoites of Neospora caninum were found in sections of lung of an equine fetus aborted 2 mo before term. Individual tachyzoites were approximately 3-5 x 2-3 microns, divided by endodyogeny, and stained positively with anti-N. caninum serum but not with anti-Toxoplasma gondii serum. Toxoplasma gondii antibody was not found in the mare's serum. This is the first report of N. caninum in a horse and indicates that N. caninum can be transmitted transplacentally in equids.     

Neospora caninum (Protozoa: apicomplexa) infections in mice

Groups of mice were given 0 mg, 4 mg, or 2 mg of methylprednisolone acetate (MPA) 7 days prior to, the day of, and 7 days after subcutaneous inoculation with 0 or 2 x 10(5) tachyzoites of Neospora caninum. Clinical signs of disease were seen only in mice given both MPA and N. caninum tachyzoites. Mice given 4 mg MPA and N. caninum tachyzoites developed severe disseminated neosporosis and most died or were killed when comatose 11-13 days postinoculation (PI). Acute pneumonia, polymyositis, encephalitis, hepatitis, and pancreatitis were the main lesions in these mice. Mice given 2 mg MPA and N. caninum developed mild pneumonia and many mice began showing neurological signs 14 days PI. Neurological signs consisted mainly of pronounced head-tilting and associated impairment of movement. Grossly visible 1-2-mm single or multiple, white areas of discoloration were seen in the brains of many of these mice. Encephalitis, ganglioradiculoneuritis, pneumonia, and polymyositis were the main changes seen in these mice. Tissue cysts of N. caninum were only seen in mice given 2 mg MPA and were first seen 21 days PI. Tissue cysts were 16-34 by 13-29 microns and had a 1.5-3.0-microns-thick cyst wall. Tissue cysts were seen only in the brain. Mice given 4 mg MPA and tachyzoites and host cells that had been frozen for 1 wk did not develop clinical signs of infection, indicating that freezing kills tachyzoites and that viruses or other agents were not involved in the genesis of disease seen in mice given MPA and viable tachyzoites.(ABSTRACT TRUNCATED AT 250 WORDS)     

Neospora caninum (Apicomplexa) in a stillborn goat.

Tissue cysts of Neospora caninum were found in sections of brain from a stillborn pygmy goat. The tissue cysts had 1-2-microns-thick cyst walls and stained with anti-N. caninum serum in an immunohistochemical test. Glial nodules, mononuclear perivascular cuffing, and foci of inflammation were associated with N. caninum tissue cysts throughout the brain. This report indicates that N. caninum can be transmitted transplacentally in goats.     

Variation of cranial and dental measurements and dental correlations in the pampean fox (Dusicyon gymnocercus)

The knowledge of the craniodental variability of South American canids is mainly anecdotal, and the variability and dental correlation of living canids have only been studied in several species of holarctic foxes ( Vulpes vulpes , Vulpes lagopus and Urocyon cinereoargeteus ) and Canis lupus . These studies have shown that the last molars, canines and incisors are the most variable and the central molars and premolars are the least variable, and that correlations are highest among the posterior premolars. This pattern has been accounted for by developmental factors, functional utility and tooth complexity. In this paper the pattern of dental correlation and variability of craniodental measurements in the South American fox Dusicyon gymnocercus is studied, with the aim of checking the extension of the pattern observed in other species and contrasting previous hypotheses. The most variable in D. gymnocercus measurements are some cranial widths, width of the incisors, lower canine and lower last molar, whereas others such as width of the braincase, length of the upper carnassials, and first and second lower and upper molars have lower variability. The premolar region is the most correlated, in terms of values and number of significant correlation coefficients. The results show that measurement error is negatively correlated with size of the variables, which could explain the bias found in this index as earlier authors have suggested, but the pattern captured by this index cannot be completely considered to be an artefact. The observed results confirm that the patterns of variability and correlation found in holarctic foxes are widespread in the family Canidae, and suggest the presence of an evolutionary constraint.     

Immune factors influencing the course of infection with Neospora caninum in the murine host

This paper investigates the role of specific immune factors on the course of infection in genetic knockout (gko) mice infected with 3 different strains of Neospora caninum. Survival time and parasite persistence were examined in interferon-gamma (IFN-gamma), tumor necrosis factor receptor-2 (TNFR2), interleukin 10 (IL-10), beta 2 microglobulin (beta2M), and inducible nitric oxide synthase (iNOS2) gko or wild-type (wt) mice following infection with either pathogenic (NC-1 or NC-2) or attenuated (NCts-8) N. caninum strains. Infection with NC-1 was 100% lethal in IFN-gamma gko mice, as evidenced by mean survival times of 10-13 days, depending on the challenge dose used. TNFR2 and beta2M gko mice infected with NC-1 or NC-2 strain demonstrated partial susceptibility to disease, as evidenced by histopathology and survival curves. TNFR2 or beta2M gko mice were not susceptible to infection with NCts-8, on the basis of absence of pathology and lack of mortality. Lack of mortality and minimal histopathology scores demonstrated that NC-1, NC-2, and NCts-8 infections were avirulent in IL-10 and iNOS2 gko mice. Adoptive transfer of immune cells from NCts-8 vaccinated normal syngeneic mice into IFN-gamma gko mice significantly (P < 0.05) prolonged mean survival times at all 3 challenge doses of NC-1 but failed to protect against mortality. Interestingly, there was a notable change in the tissue tropism of tachyzoites from the lung and brain in immunocompetent wt, TNFR2 gko, IL-10 gko, beta2M gko, and iNOS2 gko mice to the liver and spleen in IFN-gamma gko mice when challenged with N. caninum. On the basis of these results in gko mice, IFN-gamma is a critical cytokine in the host response against acute neosporosis.     

Coyotes (Canis latrans) are definitive hosts of Neospora caninum

Four captive-raised coyote pups consumed tissues from Neospora caninum-infected calves. Faeces were examined from 4 days before to 28 days after infection. One pup shed N. caninum-like oocysts, which tested positive for N. caninum and negative for Hammondia heydorni using PCR tests. Coyotes are the second discovered definitive host of N. caninum, after dogs. In North America, the expanding coyote ranges and population increase the probability of contact with domestic livestock. To reduce the risk of transmission of N. caninum to intensively farmed cattle, we recommend protection of feedstuffs using canid-proof fences, and careful disposal of dead stock.     

Plant Resources Foraged by Polybia Scutellaris (Hym. Vespidae) in the Argentine Pampas

The plants foraged by Polybia scutellaris were studied by analysing the pollen of honey collected from six nests. 34 morphological types were identified. These honeys showed a predominance of Poaceae and Cyperaceae pollen. The main access to anemophilous pollen is through the honeydew produced by Claviceps in Paspalum dilatatum. The principal sources of nectar were the native “Astereae” (Aster squamatus, Baccharis sp. and Solidago chilensis) and Eucalyptus sp. When collecting honeydew and nectar, wasps foraged plants close to their nest. Most of the plants visited belonged to a dense population. The successive flowering reflected in these honeys, shows that wasps also consume honey in summer and that it is not all stored for the winter season.     

Soil macrofauna diversity as a key element for building sustainable agriculture in Argentine Pampas

The agricultural activity in the Argentine Pampas, characterized by an important trend towards no-till soybean monocropping, has completely transformed the original Pampas landscape into a monotonous scenario with a continuous succession of farms of very low crop diversity. This process has led to soil physical, chemical and biological degradation in those systems. The increase of crop rotation rates in no-till and reduced tillage systems has been proposed as an alternative with reduced negative impact on soils in the context of conventional agriculture. On the other hand, extensive organic farming is also suggested as an alternative to high-input agriculture systems. In this article, we aim to explore how different variations of farming practices and systems impact soil macrofauna, along an edaphoclimatic gradient in the Pampas region. We studied the following systems: natural grassland (Gr) as indicator of the original community, extensive organic farming (Org), conventional agriculture with no-tillage and three crop rotation levels (Nt-R1, Nt-R2 and Nt-R3), and reduced tillage with two levels of crop rotation (Til and Til-R). We assessed soil macrofauna, with emphasis on earthworm, beetle and ant communities; and soil physical and chemical properties. Macrofaunal taxa composition was significantly affected by both management systems and edaphoclimatic conditions. The Gr community had pronounced differences from all the agricultural systems. The earthworm community from Gr had distinctive features from those of most agricultural systems, with Org and Nt-R3 being the most similar to Gr in native and exotic earthworm species, respectively. The beetle community in Org was the most different one, and the communities from the other systems did not show a pattern related to management. Ant community composition was not determined by management systems, but it was affected by edaphoclimatic conditions. All the studied macrofauna groups had a significant co-variation with soil physical and chemical properties, showing that both the characteristics of each soil relative to the geographic location and the effect of management on abiotic soil attributes have an important effect on soil macrofauna. This study confirms that biodiversity is being lost in Pampas soils, which implies a possible threat to the soil capacity to perform the processes that sustain soil functioning and hence plant productivity. Further considerations about the sustainability of the current agricultural model applied in the Argentine Pampas are needed.     

Serological evidence for naturally occurring transmission of Neospora caninum among foxes (Vulpes vulpes)

The study describes the time course of the Neospora caninum-specific antibody response in experimentally infected foxes, in naturally N. caninum-seropositive vixens and their litters. An immunofluorescence test, a tachyzoite surface antigen based ELISA and an immunoblot assay were established for this purpose. The immunoblot patterns of naturally seropositive and experimentally infected foxes revealed a high degree of similarity and resembled those reported for other intermediate host species. Reactions against immunodominant antigens of Mr 56, 68 and >94 kDa were observed which could be linked with a period of 14 days-2 months post experimental infection with tachyzoites. Cubs born by naturally seropositive vixens were found to be persistently or transiently seropositive, in the latter case, specific antibodies were detected only up to 44 days after birth. These antibodies may thus be of maternal origin. Differences between the immunoblot patterns of persistently positive cubs, those of their mothers and of transiently positive cubs, in particular the differential response to antigens of Mr 56 and 68 kDa, prove that cubs with persistent antibodies had actively mounted an antibody response. This result provides the first evidence for the postnatal or vertical transmission of N. caninum among naturally seropositive foxes.     

Status of the alien pathogen Paranosema locustae (Microsporidia) in grasshoppers (Orthoptera: Acridoidea) of the Argentine Pampas

After experimental introductions from North America in 1978–1982, the biocontrol agent Paranosema locustae became established in grasshopper communities of the western Pampas region of Argentina. The use and establishment of P. locustae in Argentina constitute both a case of neoclassical or new association biological control (use of an alien species against native pests) and a case of pathogen pollution (anthropogenic introduction–establishment of an infectious disease in populations of native species). Since P. locustae is a multihost pathogen among grasshoppers, its presence in the western Pampas represents an additional factor disrupting grasshopper communities according to the differential susceptibility of each species and possibly threatening some species. Microscopic examination of 504 grasshopper samples (the mean number of individuals per sample = 185) belonging to 43 species from 93 localities throughout the Pampas revealed an establishment area of approximate 90,000 km² from about 35° North to 38° South and from 61° East to 65° West. Field infections by P. locustae have now been detected in 21 grasshopper species in the western Pampas. Susceptible species with geographic distributions mostly restricted to the establishment area and with numerically small populations, like the melanopline Scotussa daguerrei, are predicted to be the ones facing higher risks of negative impacts.     

In vitro development of Neospora caninum (Protozoa: Apicomplexa) from dogs

The development of Neospora caninum isolated from naturally infected dogs was examined in mammalian cell cultures. Tachyzoites developed by endodyogeny when inoculated onto bovine monocyte or bovine cardiopulmonary artery endothelial cell cultures. Tachyzoites were 5.0 by 2.0 microns and had a posteriorly located nucleus. Cytopathogenic effects of parasite development consisted of the formation of holes in the cell monolayer associated with the rupture of infected host cells. Serial passage of tachyzoites was achieved by subinoculation of tachyzoites onto non-infected bovine monocyte cell cultures. It appears that N. caninum can be continuously grown in cell cultures.     

The camel (Camelus dromedarius) as an intermediate host for Hammondia heydorni

Dogs fed raw camel meat containing two types of cysts shed unsporulated Hammondia heydorni oocysts and later sporulated Sarcocystis sporocysts in their feces, but were resistant to reinfection with the Hammondia cysts. Sporulated H. heydorni occysts did not induce an enteroepithelial cycle in dogs, but resulted in the formation of muscle cysts.     

A structural study of the Neospora caninum oocyst

Oocysts of Neospora caninum were collected from the faeces of a dog fed mouse brains containing tissue cysts of the NC-beef strain of N. caninum. Sporulated oocysts were spherical to subspherical, and were 11.7x11.3 microm. The length/width ratio was 1.04. No micropyle or oocyst residuum was present. Polar granules were not present, although occasionally tiny refractile granules were seen among sporocysts. Sporocysts were ellipsoidal, did not contain a Stieda body, and were 8.4x6.1 microm. The length/width ratio for sporocysts was 1.37. A spherical or subspherical sporocyst residuum was present, and was composed of a cluster of small, compact granules of 4.3x3.9 microm, or was represented by many dispersed granules of similar size. Sporozoites were elongate and 7.0-8.0x2.0-3.0 microm in situ. No refractile bodies were present and the nucleus was centrally or slightly posteriorly located. The features of the oocyst of N. caninum are similar to those of Hammondia heydorni oocysts from dog faeces and Toxoplasma gondii and Hammondia hammondi oocysts from cat faeces.     

Evaluation of Neospora caninum serodiagnostic antigens for bovine neosporosis

Abortion and reproductive failure caused by Neospora caninum infection has a dramatic negative economic impact on the cattle industry. To date, no definitive serodiagnostic tool for assessing N. caninum abortion has been reported. In this study, we evaluated the diagnostic performance of numerous N. caninum antigens in relation to abortion in cattle. Five recombinant proteins with potential as diagnostic antigens (NcGRA6, NcGRA7, NcGRA14, NcCyP, and NcSAG1) were compared by indirect enzyme-linked immunosorbent assay (iELISA) using sera from mice and cattle experimentally infected with N. caninum. The best-performing three antigens (NcSAG1, NcGRA7, and NcGRA6) were evaluated by IgG-iELISAs to assess their utility in diagnosing Neospora abortion using sera from confirmed N. caninum-aborted dams based on immunohistochemical assays (IHC). Additionally, all samples were tested using a commercial N. caninum antibody competitive ELISA (cELISA). The iELISAs against both NcSAG1 and NcGRA7 could efficiently distinguish IHC positive and negative samples compared with iELISAs against NcGRA6 and the cELISA. Furthermore, antibody levels against NcSAG1 and NcGRA7 were significantly higher in aborting cows comparing with infected but non-aborted dams in a herd experiencing a Neospora abortion outbreak. Tracking the dynamics of antibody levels during pregnancy revealed a marked increase in NcSAG1- and NcGRA7-specific antibodies at the last trimester of pregnancy. In contrast, no marked differences in antibody levels against either antigen were noted in neurologically symptomatic calves compared with non-symptomatic infected calves. Our data suggests NcSAG1 and NcGRA7 as indicators for Neospora abortion.     

High prevalence of antibodies to Neospora caninum in white-tailed deer (Odocoileus virginianus)

Serum samples of 400 white-tailed deer (Odocoileus virginianus) from 16 preserves in northeastern Illinois were tested for Neospora caninum antibodies in the N. caninum agglutination test using mouse-derived N. caninum tachyzoites and mercaptoethanol. Antibodies were found in 162 deer with titres of 1:40 (47 deer), 1:80 (32 deer), 1:160 (17 deer), 1:200 (eight deer), 1:400 (19 deer), 1:800 (17 deer) and > or = 1:1600 (22 deer). There were no significant differences in prevalence between age or sex of the deer. The high prevalence of N. caninum infection in deer is consistent with a sylvatic cycle of N. caninum.     

Physical characterisation of the plastid DNA in Neospora caninum

The physical characteristics of the plastid DNA in Neospora caninum were investigated using pulsed-field gel electrophoresis and TEM. In a comparison of contour-clamped homogenous electric field and field inversion gel electrophoresis, the latter proved the more successful technique for studying the plastid molecules. In most cases, restriction or modifying enzymes were required to enable the plastid DNA molecules to enter the gel from the well area. The unit length of the plastid of N. caninum is approximately 35 kb; however, there is evidence for the formation of oligomeric molecules, which may migrate as linear molecules in approximate multiples of the unit length. Four different plastid genes encoding the ssrRNA, lsrRNA, rpoC and tufA genes were identified by hybridisation studies of contour-clamped homogenous electric field and field inversion gel electrophoresis gels. Transmission EM was performed on isolated plastid DNA, and circular structures similar in size and appearance to those described in other apicomplexans were observed, with an approximate length of 19 microm. The data presented here conclusively show that the Nc-Liverpool canine strain of N. caninum possesses a plastid DNA, with physical characteristics similar to the plastids found in other apicomplexans.