The 70 kDa heat-shock protein (Hsp70) sequences are considered one of the most conserved proteins in all domains of life from Archaea to eukaryotes. Hammondia heydorni, H. hammondi, Toxoplasma ...gondii, Neospora hughesi and N. caninum (Hammondia-like organisms) are closely related tissue cyst-forming coccidians that belong to the subfamily Toxoplasmatinae. The phylogenetic reconstruction using cytoplasmic Hsp70 coding genes of Hammondia-like organisms revealed the genetic sequences of T. gondii, Neospora spp. and H. heydorni to possess similar levels of evolutionary distance. In addition, at least 2 distinct genetic groups could be recognized among the H. heydorni isolates. Such results are in agreement with those obtained with internal transcribed spacer-1 rDNA (ITS-1) sequences. In order to compare the nucleotide diversity among different taxonomic levels within Apicomplexa, Hsp70 coding sequences of the following apicomplexan organisms were included in this study: Cryptosporidium, Theileria, Babesia, Plasmodium and Cyclospora. Such analysis revealed the Hammondia-like organism to be the lowest divergent group when compared to other groups within the phylum Apicomplexa. In conclusion, the Hsp70 coding sequences proved to be a valuable genetic marker for phylogenetic reconstruction and may constitute a good candidate to be used with other genes for species phylogeny within this group of organisms.
Objectives were to evaluate the effect of feeding rumen-protected methionine (RPM) in pre- and postpartum total mix ration (TMR) on lactation performance and plasma AA concentrations in dairy cows. A ...total of 470 multiparous Holstein cows 235 cows at University of Wisconsin (UW) and 235 cows at Cornell University (CU) were enrolled approximately 4 wk before parturition, housed in close-up dry cow and replicated lactation pens. Pens were randomly assigned to treatment diets (pre- and postpartum, respectively): UW control (CON) diet = 2.30 and 2.09% of Met as percentage of metabolizable protein (MP) and RPM diet = 2.83 and 2.58% of Met as MP; CU CON = 2.22 and 2.19% of Met as percentage of MP, and CU RPM = 2.85 and 2.65% of Met as percentage of MP. Treatments were evaluated until 112 ± 3 d in milk (DIM). Milk yield was recorded daily. Milk samples were collected at wk 1 and 2 of lactation, and then every other week, and analyzed for milk composition. For lactation pens, dry matter intake (DMI) was recorded daily. Body weight and body condition score were determined from 4 ± 3 DIM and parturition until 39 ± 3 and 49 DIM, respectively. Plasma AA concentrations were evaluated within 3 h after feeding during the periparturient period d −7 (±4), 0, 7 (±1), 14 (±1), and 21 (±1); n = 225. In addition, plasma AA concentrations were evaluated (every 3 h for 24 h) after feeding in cows at 76 ± 8 DIM (n = 16) and within 3 h after feeding in cows at 80 ± 3 DIM (n = 72). The RPM treatment had no effect on DMI (27.9 vs. 28.0 kg/d) or milk yield (48.7 vs. 49.2 kg/d) for RPM and CON, respectively. Cows fed the RPM treatment had increased milk protein concentration (3.07 vs. 2.95%) and yield (1.48 vs. 1.43 kg/d), and milk fat concentration (3.87 vs. 3.77%), although milk fat yield did not differ. Plasma Met concentrations tended to be greater for cows fed RPM at 7 d before parturition (25.9 vs. 22.9 µM), did not differ at parturition (22.0 vs. 20.4 µM), and were increased on d 7 (31.0 vs. 21.2 µM) and remained greater with consistent concentrations until d 21 postpartum (d 14: 30.5 vs. 19.0 µM; d 21: 31.0 vs. 17.8 µM). However, feeding RPM decreased Leu, Val, Asn, and Ser (d 7, 14, and 21) and Tyr (d 14). At a later stage in lactation, plasma Met was increased for RPM cows (34.4 vs. 16.7 µM) consistently throughout the day, with no changes in other AA. Substantial variation was detected for plasma Met concentration (range: RPM = 8.9–63.3 µM; CON = 7.8–28.8 µM) among cows coefficient of variation (CV) > 28% and within cow during the day (CV: 10.5–27.1%). In conclusion, feeding RPM increased plasma Met concentration and improved lactation performance via increased milk protein production.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii ...in 152 free-range chickens (Gallus domesticus) from 22 municipalities in 7 northeastern states (Pernambuco, Rio Grande do Norte, Maranhão, Bahia, Ceará, Sergipe, and Alagoas) of Brazil was determined. Antibodies to T. gondii were assayed by the modified agglutination test (MAT); 81 (53.3 %) chickens had titers of 1:5 in 26, 1:10 in 9, 1:20 in 4, 1:40 in 1, 1:80 in 6, 1:160 in 6, 1:320 in 13, 1:640 in 6, 1:1,280 in 3, 1:2,560 in 6, and 1:5,120 or higher in 1. Hearts and brains of 81 seropositive chickens were bioassayed individually in mice. Toxoplasma gondii was isolated from 23 chickens with MAT titers of 1:5 or higher; the isolates were designated TgCKBr165-187. Five isolates killed all infected mice. Results indicate widespread contamination of rural environment in Brazil with T. gondii oocysts.
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Attempts were made to isolate
Neospora caninum from naturally infected water buffaloes (
Bubalus bubalis) from Brazil. Brains from six buffaloes with indirect fluorescent antibodies (>1:100) to
N. ...caninum were used to isolate the parasite by bioassay in dogs and gerbils followed by in vitro culture. Shedding of
Neospora-like oocysts was noticed in dogs fed brains from three buffaloes (isolate designation NcBrBuf-1, 2 and 4). Two more isolates (NcBrBuf-3 and 5) were obtained by in vitro culture of the brains of gerbils previously infected with brains of two other buffaloes. The identity of the isolates was confirmed by biological and molecular methods. The isolates were found to be non-pathogenic to gerbils. All five isolates amplified the gene 5 amplicons using
Neospora-specific PCR assay. The sequences of gene 5 fragments and the common toxoplasmatiid ITS-1 fragments were analyzed. The dynamics of oocyst production in the dogs indicate that water buffaloes are natural intermediate hosts for
N. caninum. This is the first report of isolation of
N. caninum from water buffaloes.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
A cross-sectional study was conducted to determine the seroprevalence of anti-
Toxoplasma gondii and anti-
Neospora caninum antibodies and to investigate the risk factors related to antibodies ...against
T. gondii and
N. caninum in dogs of the city of Campina Grande, state of Paraíba, Northeast region of Brazil. For this purpose, 286 blood samples were collected from dogs during the rabies vaccination campaign, in September 2003, and on this occasion questionnaires addressing epidemiological aspects of the infections were given to each dog owner. The sera were analyzed for anti-
T. gondii and anti-
N. caninum antibodies by indirect fluorescent antibody tests. Of the total of 286 dogs, 129 were positive for
T. gondii (titer
⩾
16) with a prevalence value of 45.1% (95% CI
=
39.24–51.07%). For
N. caninum, 24 dogs were positive (titer
⩾
50), with a prevalence value of 8.4% (95% CI
=
5.45–12.23%). Antibodies to
T. gondii and
N. caninum were found simultaneously in 14 dogs (4.9%; 95% CI
=
2.7–8.08%). For
T. gondii infection, the risk factors associated with seroprevalence was the age of the animals, with dogs older than one year presenting higher values of odds ratio, and co-habitation of cats in the household. For
N. caninum infection, dogs that have street contact had higher odds of seropositivity than dogs that remained exclusively in a domestic environment.
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To determine the prevalence of
Neospora caninum antibodies and associated factors, blood sera from 623 female dairy cattle from 23 farms in the north of the state of Paraná, Brazil, were analyzed by ...means of the indirect immunofluorescent-antibody test (IFAT ≥ 25). Serum samples from 134 dogs living on the same farms also were tested for
N. caninum antibodies (IFAT ≥ 50), and the presence of dogs was associated with the prevalence of
N. caninum antibodies in cattle. The overall seroprevalence in cattle was 14.3%, mainly in animals over 24 months of age. Seroprevalence in Holsteins (15.1% of 558) was greater than in mixed-breed cattle (7.7% of 65). Age (≥24 months) of cattle, feeding silage and/or concentrate produced on the farm were associated; antibodies were found in 21.6% of dogs; and the presence of dogs was associated with the prevalence of
N. caninum antibodies in cattle.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
Antibodies to Neospora caninum and Toxoplasma gondii were assayed in sera of 396 opossums (Didelphis marsupialis) from the city of São Paulo, Brazil. Antibodies to N. caninum were assayed using the ...indirect immunofluorescent antibody test (IFAT). Antibodies (IFAT, ∼1:25) to N. caninum were found in 84 opossums (D. marsupialis) in titers of 1:25 in 46, 1:50 in 20, 1:100 in 17, and 1:400 in 1. Antibodies to T. gondii were assayed with the modified agglutination test (MAT) and the IFAT. Antibodies to T. gondii (MAT, ∼1:25) were found in 82 (20.4%) of the 396 opossums, in titers of 1:25 in 24, 1:50 in 26, 1:100 in 18, 1:200 in 13, and 1:800 in 1. The IFAT antibodies to T. gondii were found in 148 of 396 opossums, in titers of 1:16 in 41, 1:32 in 23, 1:64 in 13, 1:128 in 6, 1:256 in 20, 1:512 in 17, 1:1,024 in 10, 1:2,048 in 10, 1:4,096 in 7, and 1:8,192 in 1. This is the first report of N. caninum and T. gondii infections in D. marsupialis.
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Antibodies to Toxoplasma gondii were assayed in sera of 63 cats and 80 pigs from 71 farms located at Rondônia State, Western Amazon, Brazil, by the modified agglutination test (MAT) and the indirect ...immunofluorescent antibody test (IFAT). Antibodies (MAT ≥ 1: 25) were found in 55 of 63 cats (87.3%) with titers of 1:25 in 2, 1:50 in 2, 1:100 in 7, 1:200 in 1, 1:400 in 2, 1:800 in 9, 1:1,600 in 6, and 1:3,200 or higher in 26 cats. By IFAT, antibodies were found in 55 cats (87.3%) with titers of 1:25 in 2, 1:50 in 1, 1:100 in 4, 1:200 in 4, 1: 400 in 1, 1:800 in 13, 1:1,600 in 12, and 1:3,200 or higher in 18 cats. In pig sera, by MAT, antibodies were found in 30 of 80 pigs (37.5%) with titers of 1:25 in 2, 1:50 in 3, 1:100 in 2, 1:200 in 8, 1:400 in 3, 1:800 in 5, 1:1,600 in 3, and 1:3,200 or higher in 4 pigs. By using the IFAT (titers ≥ 1:64), antibodies were found in 35 (43.7%) pigs. The ingestion of undercooked tissues of infected pigs can be a source of T. gondii infection for humans and cats. However, the high seroprevalence of T. gondii in cats from the Amazon seems most likely to be indicative of high contamination of the environment by oocysts.
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Mechanisms of bovine corpus luteum (CL) maintenance during the second month of pregnancy have not been adequately investigated, despite significant reproductive losses. In the first month, ...interferon-tau is believed to suppress oxytocin-stimulated prostaglandin F2α (PGF) production, yet there are conflicting reports of circulating PGF metabolite (PGFM). In this study, characterization of PGFM and P4 occurred through continuous bihourly blood sampling in cows undergoing CL regression (day 18–21, n = 5), and during the first (day 18–21, n = 5) and second month (day 47–61; n = 16) of pregnancy. Cattle in the second month were assigned to control (n = 8) or oxytocin treatment (n = 8; three pulses to mimic luteolysis) to evaluate if oxytocin receptors were active. All cows but one (which had elevated PGFM prior to oxytocin treatment) maintained the pregnancy. Basal PGFM concentrations were low (11.6 ± 0.7 pg/mL) in the first month but increased 2.54-fold in the second month. Few (0.26 ± 0.12 pulses/day) PGFM pulses with low peak concentrations (28.8 ± 3.1 pg/mL) were observed during the first month of pregnancy, similar to cows not undergoing regression. However, in the second month, frequency (1.10 ± 0.26 pulses/day) and peak concentration (67.2 ± 5.0 pg/mL) of PGFM pulses increased, displaying similar frequency but lower peak PGFM than seen in regression (1.44 ± 0.14 pulses/day; 134.5 ± 18.9 pg/mL). Oxytocin treatment increased likelihood of PGFM pulses post-treatment and increased peak concentration (89.7 ± 10.1 pg/mL) in cows during the second month. Thus, cows have more PGFM pulses during second than first month of pregnancy, possibly induced by endogenous oxytocin, indicating suppression of PGF production is an important mechanism for CL maintenance during first but not second month of pregnancy. Summary Sentence Low basal PGFM concentration and minimal pulsatile activity are observed in first month of pregnancy before increasing during second month of pregnancy, suggesting different mechanisms maintain the CL in the first vs second month of pregnancy.
Background and aims
: To assess, in patients with Type 1 diabetes (T1 DM), the effects of adding a carbohydrate counting programme (CCP) to continuing education by Group Care on coping ability, ...quality of life (QoL), knowledge of diabetes, and metabolic control.
Materials and methods
: Out of 56 patients with T1 DM followed by Group Care, 27 were randomized to receive an 8-session CCP and 29 controls continued Group Care without a CCP. QoL, knowledge, and coping ability were assessed at baseline and after 30 months. Glycated hemoglobin (HbA
1c
), body weight, blood glucose, hypoglycemic episodes, and insulin dosages were checked every 3 months.
Results
: QoL improved (
p
<0.0001) in both CCP (88.7±9.2 vs78.0±9.9) and control patients (88.7±12.5
vs
80.4±11.7). At the end of study, patients on CCP had better scores in knowledge difference 0.72 (95% CI 0.44; 0.99),
p
<0.0001 and the 3 coping areas problem solving: 1.75 (1.2; 2.3),
p
<0.0001; social support seeking: −1.4 (−2.3; −0.48)
p
<0.005; avoidance: −1.59 (−2.6; −0.56),
p
<0.005 than controls. All variables showed a greater, although not statistically significant, improvement in patients with poor schooling. At 30 months, HbA
1c
was lower in the CCP patients than controls (7.2±0.9
vs
7.9±1.4),
p
<0.05. There were no changes in insulin dosage, hypoglycemic episodes or blood lipids.
Conclusions
: This study confirms that Group Care improves QoL in people with T1 DM, but suggests that specific educational and psychological supports are needed to modify adaptation to the disease. The CCP we developed appears effective in promoting change, also in patients with poor schooling.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
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