Transmission and genetic diversity of Enterococcus faecalis among layer chickens during hatch

Background

Studies on transmission of Enterococcus faecalis among chickens during hatch have not been carried out so far. Information about vertical transmission and subsequent spreading and colonization of the cloacal mucosa through cloacal ''drinking'' during hatch are important to understand the epidemiology of E. faecalis infections. In the present investigation vertical transmission and subsequent spreading and colonization of the cloacal mucosa of chickens by E. faecalis through cloacal ''drinking'' were examined.

Methods

Two different batches of layer chickens originating from 45 weeks old Brown and White Lohmann parents, respectively from the same farm were sampled in the hatcher. Isolates were confirmed to be E. faecalis by polymerase chain reaction (PCR) and further by multilocus sequence typing (MLST) to state their population structure and comparison made to sequence types previously obtained from chicken.

Results

A total of 480 chickens were swabbed from the cloacae just after hatch and after 24 hours. A total of 101 isolates were confirmed as E. faecalis by a species specific PCR. The prevalence of E. faecalis increased from 14% at 0 h to 97% after 24 h for the Brown Lohmann chickens and from 0.5% to 23% for the White Lohmann flock. The 84 isolates analysed by MLST were distributed on 14 sequence types (ST). Three ST (401, 82 and 249) accounted for 64% of all isolates analysed by MLST after 24 h. ST 82 has previously been reported from amyloid arthropathy and other lesions in poultry.

Conclusions

The present findings demonstrated a high potential of a few contaminated eggs or embryos to rapidly facilitate the spread of E. faecalis to almost all chickens during hatch.     

Prevalence of gastrointestinal nematodes in growing pigs in Kabale District in Uganda

During the last 30 years, pig production in Uganda and neighbouring counties has increased markedly. Pigs are mainly kept as a source of income for small-scale farmers; however, the pig production is subject to several constraints, one of them being worm infections. A study was carried out in rural communities in Kabale District in the South Western part of Uganda in September and October 2007 in order to estimate the prevalence of gastrointestinal nematode parasites in pigs based on coprological examination. Fifty-six households were randomly selected and visited. Housing system and deworming history were recorded. Faeces was sampled from rectum of one to five pigs (age, 3–12 months) per household. A total of 106 pigs were examined coprologically of which 91% excreted nematode eggs. The following prevalences of nematode eggs were recorded: strongyles (89%), Ascaris suum (40%), Trichuris suis (17%) and spiruroid eggs (48%). On household level, rearing pigs on slatted floors in pens significantly reduced the faecal egg excretion of strongyle eggs with almost 80% (p = 0.010) and a significant interaction between floor type and anthelmintic treatment was found for spiruroids (p = 0.037). Fifteen T. suis egg positive pigs were selected for post-mortem examination of the gastrointestinal tract. The post-mortem examinations revealed that 93% pigs were infected with Oesophagostomum spp. (worm burden, min–max 10–2,180), 73% with A. suum (1–36), 67% with T. suis (6–58), and 20% with Hyostrongylus rubidus (worms not quantified). In general, nematode infections were widespread and polyparasitism common in pigs in Uganda. However, worm burdens were moderate which may be related to recent deworming or to the practice of rearing pigs on slatted floors in wooden elevated pens.     

Selection of CMY-2 producing Escherichia coli in the faecal flora of dogs treated with cephalexin

Cephalexin is a first generation cephalosporin commonly used in dogs for treatment of pyoderma. The objective of this study was to evaluate the in vivo effects of cephalexin on selection of Escherichia coli resistant to extended-spectrum cephalosporins. A cohort study was conducted on 13 dogs presenting clinical signs of pyoderma and treated with cephalexin and 22 healthy dogs that had not been treated with antibiotics during the previous six months. Selective plating of faeces on MacConkey agar plates containing cefotaxime (CTX) yielded growth of CTX-resistant E. coli for eight of the 13 treated dogs (62%), whereas no growth was observed for any of the control dogs (Fisher exact test, P<0.001). PCR and sequence analysis identified bla(CMY-2) in all eight dogs. PCR-based replicon typing and restriction fragment length polymorphism (RFLP) of E. coli transformants revealed location of bla(CMY-2) on indistinguishable IncI1 plasmids in five of the eight dogs. One representative of these five epidemiologically related IncI1 plasmids was further characterized as sequence type (ST2) by plasmid multilocus sequence typing (pMLST). E. coli from the remaining three dogs harboured bla(CMY-2) on distinct plasmids with non-typeable replicons. A single isolate was classified as an extraintestinal pathogenic E. coli (ExPEC) due to the presence of iutA, papC and sfa/foc. The results provide a strong indication that cephalexin selects for E. coli producing plasmid-borne CMY-2 β-lactamase. The isolation of a specific IncI1 plasmid carrying bla(CMY-2) from five epidemiologically unrelated dogs suggests that cephalexin use may contribute to the spread of this plasmid lineage among Danish dogs.     

pH-induced shift in hemoglobin spectra: a spectrophotometeric comparison of atlantic cod ( Gadus morhua ) and mammalian hemoglobin

Due to a pH-sensitive effect in many fish hemoglobins (Hb), analytical errors may occur when mammalian Hb is used as a standard in quantitative spectrophotometric multicomponent analysis of fish blood. The aim of this work was to examine differences in the optical spectra of mammalian (human) and fish (farmed Atlantic cod) Hb subjected to pH 7.4 and 6.5. The absorption spectra of the common derivatives, deoxy- (HHb), oxy- (OHb), carboxy- (COHb), and methemoglobin (metHb), were determined in the spectral range of 450-700 nm. The metHb spectra of fish differed considerably from the corresponding human Hb spectra, whereas only minor differences in OHb, HHb, and COHb were found. Cod Hb was significantly (P < 0.05) influenced by a drop in pH compared to mammalian Hb. This resulted in deoxygenation of the Hb and increased autoxidation. For human Hb, a pH-independent isosbestic point in the spectra of OHb, HHb, and metHb at 523 nm was found. This isosbestic point was not found in the absorption spectra of cod Hb. In conclusion, spectra of cod metHb and human metHb behave differently. This must thus be taken into account in spectrophotometric multicomponent analysis. Ideally, Hb in muscle or blood should be determined by comparison to a standard made from the same species.     

FeLV-induced immunosuppression through alterations in signal transduction: changes in intracellular free calcium levels

Feline leukemia virus (FeLV) is a retrovirus with immunosuppressive properties. The mechanism(s) of immunosuppression is unknown. Calcium has been shown to be a second messenger in cellular activation and regulation. This study was designed to determine whether FeLV alters intracellular free calcium (IFC) levels in an FeLV-infected feline lymphoid cell line. Control cells and FeLV-infected cells were exposed to Concanavalin A, formyl-L-methionyl-L-leucyl-L-phenylalanine, and leukotriene B4. The basal IFC and post-stimulation IFC levels were recorded using Fura 2 AM and a luminescence spectrometer. Data collected indicate that FeLV-infected cells have a higher basal level of IFC and a reduced amount of increase in IFC after stimulation when compared to the control cells. The results would seem to indicate retrovirus-mediated interference occurring in the intracellular calcium signaling process.     

FeLV-induced immunosuppression through alterations in signal transduction: down regulation of protein kinase C

Activation of protein kinase C by a phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), was shown to stimulate the respiratory burst in normal cat neutrophils. Neutrophils from feline leukemia virus (FeLV)-exposed viremic and nonviremic cats had significant suppression of their respiratory burst when stimulated with TPA. The addition of whole ultraviolet light-inactivated FeLV and FeLV proteins to normal cat neutrophils produced no significant suppression of the respiratory burst. These data suggest two possible mechanisms for suppression. The first is partially due to viral alterations of the neutrophil as seen in viremic cats, but, because exogenously applied FeLV or FeLV proteins had no effect on the respiratory burst, an additional mechanism is present. The second mechanism may be caused by a latent FeLV infection residing in nonviremic cat bone marrows which alters their immune system, resulting in immunosuppression.     

A morphological study of the gross and light microscopic lesions of infectious anaemia in Atlantic salmon (Salmo salar).

Infectious anaemia in Atlantic salmon (Salmo salar) was studied by recording gross and light microscopic changes, and the development of lesions was studied in relation to haematocrit values. Gross lesions were characterised by ascites formation, congestion and enlargement of liver and spleen, congestion of the foregut and petechiae in the peritoneum. Histologically, lesions were demonstrated in the liver, being characterised by congestion in early stages (that is, haematocrit values around 25), dilatation of the sinusoids, and in later stages (haematocrit values 25 to 15) formation of blood-filled spaces bearing morphological resemblance to peliosis hepatis. At low haematocrit values (around 10), these changes comprised large areas of the liver parenchyma, that is, blood-filled areas coalesced, presenting islets of degenerate and necrotic hepatocytes. At this stage, haemorrhagic necroses were found. Spleen and kidney lesions were characterised by congestion. In the foregut, congestion and bleeding in lamina propria were observed. Liver lesions became more disseminated and severe with decreasing haematocrit values. Hypoxia due to anaemia alone cannot fully explain the development of the liver lesions.     

Immunosuppression by canine distemper virus: modulation of in vitro immunoglobulin synthesis, interleukin release and prostaglandin E2 production

In vitro or in vivo infection of canine mononuclear cells by canine distemper virus (CDV) in short-term microcultures resulted in suppression of lectin-induced 3H-thymidine incorporation. This suppressive effect was also evident in pokeweed mitogen-driven in vitro immunoglobulin synthesis and release. Lectin-induced interleukin-2 production by monocyte-depleted lymphocyte cultures was marginally affected by CDV, whereas interleukin-1 production by adherent mononuclear cells was significantly depressed. Monocyte cultures established from viremic dogs released prostaglandin (PG)E2. The results suggest that, in addition to a direct viral effect upon lectin responsive cellular population(s), CDV modulates monocyte functions by inhibition of interleukin-1 production and by enhancing PGE2 release.     

Growth potential of twelve Acacia species on acid soils in Hawaii

Reforestation of degraded tropical sites is often hampered by soils of high acidity, high aluminum saturation, and low fertility. To evaluate the possibility of cultivating Acacia species on such soils, a study was conducted at Waiawa, HI, to test growth under conditions of (1) high acidity (primarily aluminum) and nutrient stress, and (2) no acidity stress and high nutrient availability. Twelve Acacia species, including the important native Hawaiian species Acacia koa, were established on a Ustic Kanhaplohumult soil. The experimental design was a split plot with two fertility treatments as the main plots and the 12 Acacia species as subplots. The treatments were: low fertility (F₀; 143 kg ha⁻¹ 14-14-14 plus micronutrients) and high fertility (F₁; 8 Mg ha⁻¹ lime, 143 kg ha⁻¹ 14-14-14 plus micronutrients, 200 kg P ha⁻¹, and 77 kg K ha⁻¹). Acacia angustissima, Acacia aulacocarpa, Acacia auriculiformis, Acacia cincinnata, Acacia crassicarpa, Acacia implexa, Acacia koa, and Acacia mangium grew significantly faster under the high fertility treatment. Three species, A. cincinnata, A. crassicarpa, and A. mangium, are recommended for planting on infertile acid soils. The volume of A. koa was increased ten-fold by the high fertility treatment. Additional study on koa's nutritional requirements is suggested in order to identify the nutrients contributing to this increased growth.