Changes in somal growth and dendritic patterns of the retinal ganglion cells in the chicks and chick embryos

Changes in the somal growth and dendritic patterns of retinal ganglion cells (RGCs) were studied in early chick embryos and post-hatching chicks by means of the retrograde axonal transport labeling with DiI. Branching patterns of the dendrites were relatively uniform on E8 (embryonic day 8) and became more complicated on E11. Variety of the branching pattern became plainly abundant after E14. On the other hand, somata of RGCs continued to grow until E14, corresponding the appearance of the central-peripheral gradient of the somal size. After E14, RGCs elaborated on the formation of the dendritic patterns as found in chick retina, and simultaneously the growth of somal sizes almost ceased.     

Quantitative analysis of cells in the ganglion cell layer of the chick retina: developmental changes in cell density and cell size

Changes in cell density and size in the ganglion cell layer (GCL) of the retina were studied in chick embryos and post-hatching chicks. The total number of cells in the GCL increased from 3.64 million at embryonic day 8 (E8) to the maximal 7.85 million at E14. After E14, the number of cells decreased to 6.08 million at post-hatching day 1 (P1) and 4.87 million at P8. Cell density in the GCL decreased unevenly according to retinal regions; cell density in the presumptive central area (pCA) of P8-chicks decreased to approximately 45% of that in E8-embryos. Densities of the nasal peripheral retina (NP) and temporal peripheral retina (TP) of P8-chicks decreased to 23 and 18% of E8-embryos, respectively. Differentiation of the central (44,000 cells/mm(2) in pCA) - peripheral (28,000 cells/mm(2) in TP) gradient in cell density was formed by E8. The presumptive dorsal area (pDA) was shaped by E11, but became obscure with age. Although ganglion cell sizes were basically uniform at E8, differentiation occurred with the appearance of larger ganglion cells after E14. Mean size of retinal ganglion cells increased 2.8-fold in the pCA and 3.8-fold in the TP between E8 and P8, accompanying a similar scale of decreases in cell densities.     

Regional specialization of ganglion cell layer of the chick retina

Specialization of the ganglion cell layer (GCL) was studied by Nissl-staining and axonal tract-tracing methods in chicks and chick embryos. The changes in the retinal area and the cell number in the GCL produced a disparity in the cell density that occurred through the two different processes, cell generation (before embryonic days 10-14, E10-14) and cell loss (after E10-14). One high-density area was found in the retinal fundus on E8 (presumptive central area, pCA) and its density decreased toward the peripheral retina. Another high-density area was found in the dorsal retina on E11 (presumptive dorsal area, pDA). Cell densities of the pCA and the pDA on E11 decreased gradually to 25-30% by P1, and after that they further decreased to 40-60% by P30. The pCA was still identified on P30, but the pDA became very obscure by this age. In contrast, ganglion cell sizes increased 5-7 times in the pCA and pDA from E8 to P30, and increased 12 times in the temporal periphery. The present study suggests that the center-peripheral gradient of cell density results from lager scale of cell genesis in the pCA, but not from lager scale of cell loss in the peripheral retina. However, obscuration of the pDA results from equalization of cell density in cellular degeneration processes.     

Morphological classification of ganglion cells in the central retina of chicks.

Classification of retinal ganglion cells (RGCs) in the chick central retina was studied by retrograde labeling of carbocyanine dye (DiI) and intracellular filling with Lucifer Yellow. Ganglion cells were divided into 4 groups, Group Ic/Is, Group IIc/IIs, Group IIIs, Group IVc, according to sizes of somal area and dendritic field and dendritic branching pattern. Group I cells had small somal area and small dendritic field. They were further divided into 2 subgroups by complexity (subgroup Ic) and simplicity (subgroup Is) of the dendritic arborization. Group II cells had medium-sized soma and dendritic field. They were also divided into subgroup IIc and IIs by the same definitions as those of subgroup Ic and Is. Group IIIs had medium-sized soma, large and simple dendritic arborization. Group IVc in which all cells had large soma, showed large and complex dendritic arborization. Cell populations of each group were 51.8% (subgroup Ic), 21.1% (subgroup Is), 6.2% (subgroup IIc), 14.6% (subgroup IIs), 4.2% (Group IIIs), and 2.1% (Group IVc). Subgroup Ic cells, which were very similar to beta-cells in the mammalian central area, represented about a half of the ganglion cell population. Cells in subgroup Is and IIs, which were not reported in the mammalian retina, were found in the chick central retina in relatively high population (35.7%). Morphological features of chick RGCs in the central retina were considered in comparison with those of other vertebrates.     

Distinctive histopathologic features of canine optic nerve hypoplasia and aplasia: a retrospective review of 13 cases

Canine optic nerve hypoplasia (ONH) and aplasia (ONA) are significant neuro-ophthalmologic disorders that have been reported in several species. The purpose of this study was to describe the distinctive histopathologic features of ONH and ONA in canine patients identified from a collection of 20 000 ocular submissions at the comparative ocular pathology laboratory of Wisconsin from 1989 to 2006. The following information about ONH and ONA cases was collected: signalment, and clinical and gross findings, including unilateral vs. bilateral involvement. Microscopic evaluation was performed, with attention to optic nerve malformation, retinal ganglion cell (RGC) and nerve fiber layer (NFL) loss, and retinal disorganization. The distribution of retinal vasculature was recorded and a search for unusual findings of ONH and ONA was performed. Information and histologic documentation was available for 13 cases. Eight cases of ONH and five cases of ONA were identified. The average group age was 20.2 months and 16.1 months, respectively. The most common breed was the Shih Tzu (3/13). ONH usually presented bilaterally (7/8); all ONA cases presented as a unilateral disease (5/5). The morphologic findings in the optic nerve (ON) in ONH included variable degrees of ON hypoplasia and gliosis, as well as ectopic vestigial ON remnants within orbital nerves and connective tissues. The NFL was detected in the majority of the ONH cases; however, RGCs were rare or absent. Mild retinal disorganization was seen occasionally. Most cases of ONH were associated with regional peripheral retinal blood vessel extension into the vitreous, leaving the peripheral retina avascular. In ONA cases the retinal blood vessels, NFL and RGCs were totally absent and retinal disorganization was severe. Distinctive microscopic features encountered in ONA included anterior segment dysgenesis in some cases. The retina in these cases was stretched across the posterior lens capsule, never making contact with the posterior pole of the globe. The current study reviews the human and veterinary literature pertaining to ONH and ONA, compares ONH and ONA in dogs, and presents related ophthalmic histopathologic findings that have not been reported previously.     

Regional specialization of the Ganglion cell density in the retina of the Ostrich (Struthio camelus)

In this study, retinal whole-mount specimens were prepared and stained with 0.1% cresyl violet for the ganglion cell study in the Ostrich (Struthio camelus) . The total number, distribution, and size of these cells were determined in different retinal regions. The mean total number of ganglion cells (three retinas) was 1 435 052 with an average density of 652 cells/mm². The temporo – nasal area of the retina with high cell density were identified with the peak of 7525 cells/mm² in the central area. The size of most ganglion cells ranged from 113–403 µm², with smaller cells predominating along the temporo-nasal streak above the optic disc and larger cells comprising more of the peripheral regions. The average thickness of the retina was 196 µm. The central area was the thickest area (268.6 µm), whereas the peripheral area was the thinnest area. Thus, the specialization of ganglion cell densities, their sizes and the thickness of the retina support the notion that the conduction of visual information towards the brain from all regions of the retina is not uniform, and suggests that the temporo – nasal streak is the fine quality area for vision in ostriches.     

Regional Specialisation of the Ganglion Cell Density in the Retina of the Native Duck (Anas platyrhynchos) of Bangladesh

In this study, retinal whole‐mount specimens were prepared and stained with 0.1% cresyl violet for the ganglion cell study in the native duck (Anas platyrhynchos). The total number, distribution and size of these cells were determined in different retinal regions. The mean total number of ganglion cells was 1 598 501. The retinal area centralis had the highest ganglion cell density with 11 200 cells/mm². Number of ganglion cell bodies was the highest in temporal area, followed by dorsal, nasal and ventral areas. Ganglion cell size ranged from 5.25 to 80 μm². In the temporal and nasal region, most of the cells were ranged from 15 to 25 μm², and in the dorsal and ventral region, most of the cells were ranged from 12 to 25 μm². There was a marked trend for the retinal ganglion cell size to increase as the population density decrease towards the periphery. A population of small ganglion cells persisted into the central area just above the optic disc and the largest soma area was in the ventral zone of the retina. Thus, the specialisation of ganglion cell densities and their sizes support the notion that the conduction of visual information towards the brain from all regions of the retina is not uniform, and the central area is the fine quality area for vision in native duck.     

Retrograde tracing with fluorescent microspheres reveals bifurcating projections from central retina to tectum and thalamus in chicks

The goal of this study is to demonstrate the dual-projection pattern of retinal ganglion cells (RGCs) projecting to the tectum and visual thalamus in chick using retrograde fluorescent tracers and also to define the morphological properties of these RGCs with dual projections by intracellular injection of Lucifer Yellow (LY) combined with immunohistochemistry. Thirty-two chicks received double injections of green and red fluorescent microspheres into their thalamus and tectum in the same side. In the central retina, most of the labelled RGCs were tec-RGCs (RGCs projecting to the tectum), a quarter was tha-RGCs (RGCs projecting to the thalamus), and almost all of the tha-RGCs were double-labelled RGCs. An intracellular injection of LY into the double-labelled RGCs showed all six groups of RGCs without specific populations in each group (J. Comp. Neurol., 2004, 469: 360). These dendritic patterns were mostly mono- and bistrata, which extended horizontally in the deeper part of the inner plexiform layer.     

Topography of ganglion cells in the retina of the duck (Anas platyrhynchos var. domesticus)

The purpose of this study was to define ganglion cell density, size and topography in the retina of the mallard duck. After killing adult mallard ducks (Anas platyrhynchos var. domesticus), their eyes were removed using pentobarbital (30 mg/kg). The retinas were isolated, whole mount specimens were prepared by staining with 0.1% cresyl violet and then fixing the tissues for study. The retinal ganglion cells were counted, mapped and measured. The mean total number of ganglion cells was estimated at approximately 1.7 × 10⁶ and the retinal area centralis had the highest ganglion cell density with 15 820 cells/mm². The number of ganglion cell bodies was highest in the temporal area, followed by the nasal, dorsal and ventral areas. Ganglion cell size ranged from 56 to 406 μm². A population of small ganglion cells persisted into the central area just above the optic disc and the largest soma area was in the ventral zone of the retina. This localization of ganglion cells suggests that the quality of vision is not equal in all the areas of the duck retina and the central part may have the highest vision quality as a function of the retinal ganglion cells.     

The neuroprotective effects of lidocaine and methylprednisolone in a rat model of retinal ischemia-reperfusion injury

Retinal ischemia is a common cause of visual impairment for humans and animals. The neuroprotective effects of lidocaine (LDC) and methylprednisolone (MP) upon retinal ischemic injury were investigated in a rat model. Sprague-Dawley rats were divided into 3 groups, the IR control, LDC and MP. A very high intraocular pressure (HIOP) and retinal ischemia were induced. In LDC group, LDC bolus (1.5 mg/kg) was i.v. injected 30 min before ischemia and then a constant rate infusion (CRI) with 2 mg/kg/hr was given until 60 min after reperfusion. In MP group, MP bolus (30 mg/kg) was i.v. administered twice at 2 min before and immediately after ischemia, respectively. The HIOP damage to retina was evaluated by electroretinogram (ERG) and morphometrical histology. The functional analysis of the retina by ERG revealed a 35.2% reduction of a-wave in the IR group, 49.7% reduction in the LDC group but no significant change in the MP group compared to normal controls. An 81.0% reduction of b-wave was observed in the IR group, 80.7% reduction in the LDC group and 17.6% reduction in the MP group. In the morphometrical histology, the retinal inner plexiform layer/outer nuclear layer (IPL/ONL) ratio was reduced to 48.8% in the IR group, 80.1% in the LDC group and 96.2% in MP group. In conclusion, the MP showed significantly good neuroprotective effects on retinal IR injury, and the LDC showed moderate neuroprotective effects demonstrated in retinal structure but not in retinal function.     

Assessment of the posterior segment of the cat eye by optical coherence tomography (OCT)

OBJECTIVES: To assess the feasibility of optical coherence tomography (OCT) for examining the cat ocular fundus, to provide normative data on retinal thickness in different fundus regions, and to demonstrate selected surgically induced vitreoretinal pathologies in the cat. ANIMAL STUDIED: Forty-five eyes of 28 healthy domestic cats and two eyes of domestic cats that had undergone subretinal implantation surgery for a visual prosthesis were examined. PROCEDURES: An optical coherence tomograph (Zeiss-Humphrey) was used to examine the anesthetized animals. At least five vertical and five horizontal scans in regular distribution were recorded for each cat including (1) the peripapillary region, (2) the area centralis, and (3) the peripheral retina. Thickness was measured manually at five locations in each scan. Retinal thickness was compared in the three above-mentioned fundus regions, between eyes and between vertical and horizontal scans. OCT was additionally performed in animals with retinal detachment and a subretinal visual prosthesis. RESULTS: OCT measurements required only minimal adjustments of human settings and yielded high quality images. In comparison to humans intraretinal layers were more difficult to differentiate. Retinal thickness was highest in the peripapillary region (245 +/- 21 microm), followed by the peripheral retina (204 +/- 11 microm) and the area centralis (182 +/- 11 microm; all P < 0.0001). There was no statistically significant difference between right and left eye or between vertical and horizontal scans. OCT demonstrated retinal detachment, an iatrogenic break and a subretinal prosthetic device in high detail. CONCLUSIONS: Retinal thickness was measurable with high precision; values compare well to older histologic studies. OCT bears significant advantages over histology in enabling one to repeat measurements in living animals and thus allowing longitudinal studies. Various vitreoretinal pathologies common in feline eyes are detectable and quantifiable by OCT.     

Number and density of retinal photoreceptor cells with emphasis on oil droplet distribution in the Mallard Duck (Anas platyrhynchos var. domesticus)

This study was intended to determine the number and regional distribution of photoreceptor cells and different colored oil droplets in the retina of the Mallard Duck (Anas platyrhynchos var. domesticus). To estimate the number and density of photoreceptor cells, adult ducks were killed and both eyes were enucleated under deep anesthesia to prepare Nissl‐stained retinal whole‐mount samples. Different colored oil droplets were counted from color microphotographs of the freshly prepared retina. The mean number of retinal photoreceptors was approximately 6 308 828 ± 521 927, with a peak density of 33 573/mm² in the central retina. The density was similar in the nasal, temporal, ventral and dorsal areas of the retina. Five types of oil droplets were identified on the basis of color: red, orange, greenish‐yellow, yellow and clear. The mean density of oil droplets was highest in the central retina (17 639/mm²) and gradually declined towards the nasal, temporal, ventral and dorsal areas. The size of oil droplets gradually increased with retinal eccentricity and varied even within an area. The greenish‐yellow oil droplets were most abundant across the retina. Taken together, these results demonstrate the differential retinal distribution of photoreceptor cells and oil droplets in duck retina. We conclude that the area of high photoreceptor cell density, which is matched by high neuron densities of the ganglion cell layer, corresponds to the site of acute vision in duck retina.     

Observations on the effects of long-term withdrawal on carcass composition and residue concentrations in clenbuterol-medicated cattle

The detection of the illegal use of clenbuterol (CBL) as a growth promoter has relied on detecting residual concentrations of the drug in body fluids or tissues. Analysis of retinal extracts has recently been shown to considerably extend the detection period following withdrawal. The withdrawal periods required to eliminate residues from the liver and retina were investigated by medicating 20 cattle with CBL for 30 days; 6 control animals remained unmedicated. Residual concentrations were monitored throughout this period and for the subsequent 140 days. Concurrent changes in muscle areas and backfat thicknesses were recorded by ultrasound.CBL was detectable in liver up to the 56th day of withdrawal (0.35 ng/g, SD=0.5), but retinal concentrations remained well above detectable concentrations throughout the withdrawal period (22.5 ng/g, SD=6.5). There were small gains (3–4%) in the muscle areas of treated cattle during medication as compared to controls (p>0.05). These comparative gains remained during withdrawal. Backfat thicknesses in treated animals were 40% lower than in controls at the end of medication (p<0.01). However, by 70 days after withdrawal this difference had disappeared (p>0.05) owing to accelerated fat deposition in the treated group. The retina has been shown to be a highly effective target matrix for detecting CBL administration after long withdrawal periods.Abbreviations CBL clenbuterol - CIM cimaterol - EC European Community - EIA enzyme immunoassay - GC-MS gas chromatography-mass spectrometry - HRPo horseradish peroxidase - L. dorsi, Longissimus dorsi - MRL maximum residue level - SD standard deviation - SED standard error of the difference     

In vivo imaging reveals novel aspects of retinal disease progression in Rs1h(-/Y) mice but no therapeutic effect of carbonic anhydrase inhibition

Purpose X-linked juvenile retinoschisis (XLRS) is the most common juvenile maculopathy in men and is caused by mutations in the gene encoding retinoschisin (RS1). Evidence in the literature on the therapeutic effect of carboanhydrase inhibitors (CAIs) to treat schisis formation in the retina has remained equivocal. Here, we evaluate the effect of the CAI dorzolamide on the structural and functional disease progression in the mouse model for XLRS (Rs1h(-/y) ). Methods Rs1h ( -/y ) mice were treated unilaterally with dorzolamide eye drops (Trusopt(?) 20?mg/mL) every 12?h for 2?weeks starting on postnatal day 14 (n?=?27). Changes of retinal structure were monitored by confocal scanning laser ophthalmoscopy and spectral domain optical coherence tomography 12?h, 14?days, 4?weeks, 2?months, and 6?months after completion of the treatment. Results Schisis formation (peak at 3?months) preceded photoreceptor degeneration and hyper-fluorescence (peak at 7?months). Structural pathology was most severe in the superior hemi-retina with previously unreported hyper-fluorescent lesions. Quantitative analysis showed no significant differences regarding the inner or outer retinal thickness of the treated vs. untreated eyes 12?h after the completion of treatment (IRT(12?h) =?-1.29?±?1.89?μm; ORT(12?h) =?0.61?±?2.08?μm; mean?±?95%CI) or at any later time point. Conclusion Time line analysis after short-term treatment with CAI failed to show short-, intermediate-, or long-term evidence of structural improvement in Rs1h(-/y) mice. Schisis formation in the inner retina peaked at the age of 3?months and was followed by photoreceptor degeneration predominantly in the superior hemi-retina. Previously unreported hyper-fluorescent lesions co-register with structural retinal pathologies.     

Age changes in the eyes of the Beagle dog

Changes with age in the eyes of Beagle dogs are described clinically and histologically. Lenticular changes involve increased prominence of the posterior lens sutures and opacity within the fetal nucleus. Cystoid degeneration of the retina was found in 85% of 8–year–old animals examined. A pigmentary change of the retina was found to be associated with focal degenerative change of the tapetum. No case of retinal atrophy was recorded.     

Effects of exogenous estradiol-17 beta on luteinizing hormone, progesterone, and estradiol-17 beta concentrations before and after prostaglandin F2 alpha-induced termination of pregnancy and pseudopregnancy in gilts.

This study evaluated the influence of exogenous estradiol-17 beta (E2) administration on LH concentrations and the number of animals returning to estrus after the termination of pregnancy or pseudopregnancy in gilts. Gilts were mated (pregnant; n = 11) on the 1st d of estrus or received 5 mg of estradiol valerate i.m. at d 11 to 15 after the onset of estrus (pseudopregnant; n = 9). Gilts were treated with prostaglandin F2 alpha (PGF2 alpha, 15 and 10 mg) at 12-h intervals on d 44 of pregnancy or pseudopregnancy. The day of abortion or luteolysis (progesterone less than .2 ng/mL) was considered d 0. Six pregnant and four pseudopregnant gilts received s.c. an E2 capsule (24 mg of E2) on d -20 and additional E2 capsules on d -13 and -6. The E2 capsules were removed on the day after PGF2 alpha administration. Blood samples were collected at 12-h intervals from d -21 to -3, at 6-h intervals from d -2 to 21 or the onset of estrus, and at 15-min intervals for 8 h on d -2, 1, 4, 7, 10, 14, and 18. After each 8-h sampling period, gilts were treated i.v. with GnRH at .5 micrograms/kg of BW and blood samples collected at 10-min intervals for 3 h. A greater (P less than .05) proportion of sham-treated gilts than of E2-treated gilts exhibited a preovulatory-like LH surge after abortion/luteolysis. It was evident that E2 supplementation before luteolysis reduced the ability of pregnant and pseudopregnant gilts to return to estrus.     

Intracapsular lensectomy and sulcus intraocular lens fixation in dogs with primary lens luxation or subluxation

Objective  To evaluate the postoperative results of lensectomy and sulcus intraocular lens fixation (SIOLF) via an ab interno approach in dogs with progressive lens subluxation or early luxation.
Study design  Retrospective study.
Animals studied  Twenty eyes from 19 dogs presented to the Animal Eye Clinic for lens luxation or subluxation between 1999 and 2006.
Methods  Medical records were reviewed to evaluate preoperative lens position, vision status, intraocular pressure (IOP), and whether surgery was performed on an emergent or elective nature. Lensectomy and SIOLF were performed and postoperative status including vision, glaucoma, and retinal detachment was assessed.
Results  Average age was 8.6 years (range 4–14 years) and 55% (11/20) were terriers. Patients were followed a mean of 29.2 months (range 1–92 months) after surgery. Retinal detachment or secondary glaucoma was observed in 1 of 20 (5%) and 5 of 20 (20%) eyes, respectively, with 1 of 20 (5%) exhibiting both. Mean preoperative IOP was 16 mmHg and preoperative lens position was equally divided between luxated and subluxated lenses. Surgery was performed more frequently as an elective procedure (18/20; 90%) due to normalized IOP vs. an emergency procedure (2/20; 10%). Vision was retained in 70% (14/20) of eyes with a mean time to vision loss of 41 months in the remaining eyes due to glaucoma, retinal detachment, or retinal degeneration.
Conclusions  Complications of glaucoma and retinal detachment after SIOLF in this study were less when compared with previously reported incidence rates in the literature for lensectomy alone which may reflect improved patient selection.     

Evidence for transuterine migration of embryos in the domestic cat

A total of 169 pregnant cats presented for ovariohysterectomy were examined for the number of corpora lutea (CL) and the number of fetuses per uterine horn. The implantation rate and the frequency of occurrence of transuterine migration of embryos were calculated. The average number of CL was 5.6 +/- 1.9 (mean +/- SD; range, 2-11), the average number of fetuses was 4.5 +/- 1.4 (range, 1-8) and the average implantation rate was 84 +/- 20%. Transuterine migration occurred in 69/169 cats (40.8%). In animals with a 100% implantation rate, transuterine migration occurred in 50/84 cats (59.5%). The number of embryos that migrated ranged from 1-3 per animal and embryos moved from the uterine horn ipsilateral to the ovary with the larger number of CL in 66/69 cats (95.7%). As a result of transuterine migration, the difference in the number of fetuses between uterine horns as compared with the difference in the number of CL between ovaries was smaller in 54 casts (78.3%), unchanged in 8 cats (11.6%) and larger in 7 cats (10.1%). These results indicate that cats tend to equalize the number of fetuses between uterine horns by transuterine migration of embryos.     

北疆奶牛隐性乳房炎的调查与分析

[目的]为了解北疆某牧场奶牛隐性乳房炎情况及分析其可能存在的风险因素。[方法]2020年1月至2022年9月通过加州乳房炎检测法（CMT）对该牧场在群泌乳牛进行隐性乳房炎调查,并初步分析该场可能存在的主要风险因素。[结果]该牧场2020—2022年各年奶牛隐性乳房炎平均阳性率分别为3.69%（110/2 980）、5.01%（150/2 993）和3.87%（86/2 220）;乳区阳性率分别为0.93%（110/11 856）、1.35%（160/11 880）和0.97%（86/8 860）;瞎乳头率分别为0.13%（16/11 920）、0.19%（23/11 972）和0.23%（20/8 880）;夏季隐性乳房炎阳性率明显高于其他季节,奶牛各乳区隐性乳房炎阳性率差异不显著（P>0.05）,不同胎次间的奶牛隐性乳房炎平均阳性率差异性显著（P<0.05）,5胎和6胎及以上牛的隐性乳房炎平均阳性率均明显高于其他胎次。[结论]该牧场奶牛隐性乳房炎发生率较低但仍需改善,夏季炎热潮湿的饲养环境和高龄奶牛更易发生热应激,奶牛身免疫力下降等原因可能是该牧场奶牛患乳房炎的主要风险因素。     

The afa-related gene cluster in necrotoxigenic and other Escherichia coli from animals belongs to the afa-8 variant

Six hundred and nine necrotoxigenic Escherichia coli type 1 and 2 (NTEC1 and NTEC2) and non-NTEC isolated in Western and Southern Europe, North Africa and Canada from diseased calves, pigs, humans, poultry, and 55 isolated from asymptomatic calves were studied for the identification of afa-related sequences to the recently described afa-7 and afa-8 gene cluster variants from two bovine Escherichia coli (Lalioui et al., 1999). Colony hybridization and PCR assays for the afaD-7, afaE-7, afaD-8 and afaE-8 identified the afa-related sequences to the afa-8 gene cluster in most (67/79; 85%) of the E. coli positive with the Afa-f family probe and in 14 additional strains negative with the Afa-f probe. No E. coli was positive for the afa-7 gene cluster. The existence of afa-8 positive strains was thus confirmed among bovine E. coli and for the first time among porcine, poultry and human E. coli. Sequencing of the afaE-8 amplicon of nine strains from the different host species showed a high degree of conservation (>95% at the DNA level; >92% at the amino-acid level). The afa-8 gene cluster was more frequent in E. coli from diseased calves (18%) than from piglets (12%), humans (6%) and poultry (5%). Bovine NTEC2 (26%) were more frequently positive than NTEC 1 (20%) and non-NTEC (11%). E. coli isolated from asymptomatic calves were rarely positive: one NTEC2 (3%) and no non-NTEC. The afa-8 gene cluster was located on the Vir plasmid in 11/23 NTEC2, but no plasmid localization was detected in NTEC1 or non-NTEC.