Experimental Neospora Caninum Infection in Pregnant Dairy Heifers Raises Concentrations of Pregnancy‐Associated Glycoproteins 1 and 2 in Foetal Fluids

Plasma concentrations of PAG‐1 are used for pregnancy diagnosis and as a marker of placental/foetal well‐being, while those of PAG‐2 may be an indicator of abortion risk in Neospora caninum‐infected cows. Studies have shown that N. caninum infection modifies PAG‐1 and PAG‐2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG‐1, PAG‐2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG‐2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non‐infected foetuses. In infected foetuses, significantly higher concentrations of PAG‐1 (p < 0.001) and PAG‐2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non‐infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG‐1 and PAG‐2 levels and pH values than fluids from non‐infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.     

Comparison of diagnostics techniques in an outbreak of infectious laryngotracheitis from meat chickens

Various diagnostics techniques were compared for their ability to detect infectious laryngotracheitis (ILT) during an outbreak in chickens aged between 4 and 21 wk. Gross lesions ranged from excess mucus to accumulation of fibrinonecrotic exudate in the larynx and trachea. Syncytial cells with intranuclear inclusion bodies were found in sinus, conjunctiva, larynx, trachea, lung, and air sac. Virus isolation in chicken embryos was attempted in every case. Negative-stain electron microscopy detected herpesvirus in only 6% of the cases. Yet, isolation of ILT virus in the chorioallantoic membrane was presumed by histology in >20% of the samples and confirmed by fluorescent antibody (FA) in 35% of the embryos inoculated with conjunctivas or tracheas from affected birds. Overall, results from histology and FA tests were highly correlated. FA test has the advantage over histology of being diagnostically specific for ILT virus. Polymerase chain reaction was the most sensitive test and detected the viral DNA even in cases where histology and FA were negative. ILT virus DNA was quantified by real-time polymerase chain reaction (Re-Ti ILTV). Histologic and FA results from larynx and trachea were negative if the concentration of the viral DNA was < or =4 of log10. A viral DNA concentration higher than log10 4, as determined by Re-Ti ILTV, was required for clinical ILT to be manifested.     

Multiple generalized follicular cysts in a stallion

This case report describes a case of multiple follicular cysts in a 4-year-old Spanish purebred stallion. The lesions ranged in size from 0.5 to 3 cm in diameter, and were firm, well circumscribed and nonpruritic. They developed over a 2-year period with a generalized distribution affecting all body regions. Five nodules were removed and histopathologically corresponded to simple epidermal cysts (infundibular and isthmus-catagen) with squamous epithelium and a keratin filled cavity. Lesions were not evident at birth but their number, early age of detection, slow growth and lack of previous trauma suggested that they were congenital. To the authors' knowledge, this condition has not previously been reported either in young horses or in Spanish purebred horses.     

Canine bartonellosis: serological and molecular prevalence in Brazil and evidence of co-infection with Bartonella henselae and Bartonella vinsonii subsp. berkhoffii

The purpose of this study was to determine the serological and molecular prevalence of Bartonella spp. infection in a sick dog population from Brazil. At the S?o Paulo State University Veterinary Teaching Hospital in Botucatu, 198 consecutive dogs with clinicopathological abnormalities consistent with tick-borne infections were sampled. Antibodies to Bartonella henselae and Bartonella vinsonii subsp. berkhoffii were detected in 2.0% (4/197) and 1.5% (3/197) of the dogs, respectively. Using 16S-23S rRNA intergenic transcribed spacer (ITS) primers, Bartonella DNA was amplified from only 1/198 blood samples. Bartonella seroreactive and/or PCR positive blood samples (n=8) were inoculated into a liquid pre-enrichment growth medium (BAPGM) and subsequently sub-inoculated onto BAPGM/blood-agar plates. PCR targeting the ITS region, pap31 and rpoB genes amplified B. henselae from the blood and/or isolates of the PCR positive dog (ITS: DQ346666; pap31 gene: DQ351240; rpoB: EF196806). B. henselae and B. vinsonii subsp. berkhoffii (pap31: DQ906160; rpoB: EF196805) co-infection was found in one of the B. vinsonii subsp. berkhoffii seroreactive dogs. We conclude that dogs in this study population were infrequently exposed to or infected with a Bartonella species. The B. henselae and B. vinsonii subsp. berkhoffii strains identified in this study are genetically similar to strains isolated from septicemic cats, dogs, coyotes and human beings from other parts of the world. To our knowledge, these isolates provide the first Brazilian DNA sequences from these Bartonella species and the first evidence of Bartonella co-infection in dogs.     

Natural hybridization between wheat (Triticum aestivum L.) and its wild relatives Aegilops geniculata Roth and Aegilops triuncialis L.

BACKGROUND

Cultivated bread wheat (Triticum aestivum L.) spontaneously hybridizes with wild/weedy related Aegilops populations, but little is known about the actual rates at which this hybridization occurs under field conditions. It is very important to provide reliable empirical data on this phenomenon in order to assess the potential crop–wild introgression, especially in the context of conducting risk assessments for the commercialization of genetically modified (GM) wheat, as gene flow from wheat to Aegilops species could transfer into the wild species genes coding for traits such as resistance to herbicides, insects, diseases or environmental stresses.

RESULTS

The spontaneous hybridization rates between wheat and A. geniculata and A. triuncialis, which are very abundant in the Mediterranean area, have been estimated for the first time in the northern part of the Meseta Central, the great central plateau which includes the largest area of wheat cultivation in Spain. Hybridization rates averaged 0.12% and 0.008% for A. geniculata and A. triuncialis, respectively. Hybrids were found in 26% of A. geniculata and 5% of A. triuncialis populations, at rates that can be ≤3.6% for A. geniculata and 0.24% for A. triuncialis.

CONCLUSION

The detection of Aegilops spp.–wheat hybrids in Aegilops populations indicates that gene flow can occur, although wheat is considered a crop with a low-to-medium risk for transgene escape. These data on field hybridization rates are essential for GM wheat risk assessment purposes. © 2023 Society of Chemical Industry.     

Genetic basis and origin of resistance to acetolactate synthase inhibitors in Amaranthus palmeri from Spain and Italy

BACKGROUND

Amaranthus palmeri is an aggressive annual weed native to the United States, which has become invasive in some European countries. Populations resistant to acetolactate synthase (ALS) inhibitors have been recorded in Spain and Italy, but the evolutionary origin of the resistance traits remains unknown. Bioassays were conducted to identify cross-resistance to ALS inhibitors and a haplotype-based genetic approach was used to elucidate the origin and distribution of resistance in both countries.

RESULTS

Amaranthus palmeri populations were resistant to thifensulfuron-methyl and imazamox, and the 574-Leu mutant ALS allele was found to be the main cause of resistance among them. In two Spanish populations, 376-Glu and 197-Thr mutant ALS alleles were also found. The haplotype analyses revealed the presence of two and four distinct 574-Leu mutant haplotypes in the Italian and Spanish populations, respectively. None was common to both countries, but some mutant haplotypes were shared between geographically close populations or between populations more than 100 km apart. Wide genetic diversity was found in two very close Spanish populations.

CONCLUSION

ALS-resistant A. palmeri populations were introduced to Italy and Spain from outside Europe. Populations from both countries have different evolutionary histories and originate from independent introduction events. ALS resistance then spread over short and long distances by seed dispersal. The higher number and genetic diversity among mutant haplotypes from the Spanish populations indicated recurrent invasions. The implementation of control tactics to limit seed dispersal and the establishment of A. palmeri is recommended in both countries. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Fertility of Holstein cows and heifers submitted to timed artificial insemination and receiving one or two doses (12 h apart) of semen

The objective of this retrospective study was to assess the effect of receiving a single (n = 50,285) or double (n = 4392) artificial insemination (AI), 12 h apart, within a timed artificial insemination protocol on pregnancy per AI (P/AI) in nulliparous heifers (inseminated with either sex-sorted or conventional semen) and pluriparous Holstein cows in a commercial dairy herd. Also, this study aimed to investigate the relationship between temperature-humidity index (THI) and time of the first AI and fertility. Fertility of cows receiving two AI with normothermia (THI <68) was higher (p < .05) than cows receiving a single AI (42.9% vs. 36.4%). P/AI of cows receiving two AI with severe heat stress (THI >85) was higher (p < .05) than cows receiving a single AI (21.0% vs. 12.6%). Regardless of heat stress conditions, applying the first AI in the morning increased (p < .05) P/AI in cows with double AI than in cows whose first AI occurred in the afternoon (38.4 vs. 33.3%). With moderate heat stress, and sexed-sorted semen, P/AI to timed AI was higher (65.0 vs. 51.9%; p < .05) in heifers receiving double AI than those serviced once. It was concluded that double AI, 12 h apart, enhanced fertility at timed AI than herd mates with a single AI, particularly with heat stress at breeding.