Studies on prolonged spermatozoa survival in chiroptera: A morphological examination of storage and clearance of intrauterine and cauda epididymal spermatozoa in the bats Myotis lucifugus and M. velifer

The cauda epididymidis, uterine corpus, and cornua and uterotubal junction of Myotis function to retain and preserve normal spermatozoa throughout hibernation. In none of the sites do spermatozoa show features that might account for their extended viability. Spermatozoa stored in the uterus and epididymis show no special orientation toward the epithelium lining these sites, whereas an intimate relationship is established between some sperm and the epithelial cells of the uterotubal junction which might either account for extended postcoital sperm survival or forecast their removal from further participation. Transmission and scanning electron microscopic observations do not disclose any morphological changes in stored luminal spermatozoa. A low rate of phagocytosis of sperm is evident in the female tract during hibernation. However, spermatozoa are evidently not vulnerable to being removed from the storage sites until spring arousal when ovulation occurs. Both uterotubal epithelial cells and phagocytes appear to be involved in the disposal of spermatozoa in the female, whereas epididymal spermatozoa apparently are primarily voided during urination. A mechanism that delays capacitation must underlie the ability of spermatozoa to survive in the female reproductive tract of the hibernating bat.     

Two novel superantigens found in both group A and group C Streptococcus

Two novel streptococcal superantigen genes (speL(Se) and speM(Se)) were identified from the Streptococcus equi genome database at the Sanger Center. Genotyping of 8 S. equi isolates and 40 Streptococcus pyogenes isolates resulted in the detection of the orthologous genes speL and speM in a restricted number of S. pyogenes isolates (15 and 5%, respectively). Surprisingly, the novel superantigen genes could not be found in any of the analyzed S. equi isolates. The results suggest that both genes are located on a mobile element that enables gene transfer between individual isolates and between streptococci from different Lancefield groups. S. equi pyrogenic exotoxin L (SPE-L(Se))/streptococcal pyrogenic exotoxin L (SPE-L) and SPE-M(Se)/SPE-M are most closely related to SMEZ, SPE-C, SPE-G, and SPE-J, but build a separate branch within this group. Recombinant SPE-L (rSPE-L) and rSPE-M were highly mitogenic for human peripheral blood lymphocytes, with half-maximum responses at 1 and 10 pg/ml, respectively. The results from competitive binding experiments suggest that both proteins bind major histocompatibility complex class II at the beta-chain, but not at the alpha-chain. The most common targets for both toxins were human Vbeta1.1 expressing T cells. Seroconversion against SPE-L and SPE-M was observed in healthy blood donors, suggesting that the toxins are expressed in vivo. Interestingly, the speL gene is highly associated with S. pyogenes M89, a serotype that is linked to acute rheumatic fever in New Zealand.     

Class II DRB polymorphism and sequence diversity in two vesper bats in the genus Myotis

Almost no studies have been done with respect to major histocompatibility complex (MHC) polymorphism and sequence diversity in bats, although they account for one in five living mammalian species. We analysed MHC Class II DRB polymorphism and sequence diversity in two Mexican verpertilionid bat species, the widespread continental species Myotis velifer and the narrowly distributed (and endangered) island endemic Myotis vivesi. We find extensive DRB polymorphism in the widespread M. velifer, similar to that commonly reported in other mammals. The geographically restricted M. vivesi by contrast shows only very limited polymorphism. We conclude that M. vivesi has undergone a dramatic loss of MHC polymorphism. The significance of this inference in light of other information on population structure and genetic diversity in this species is discussed.     

Ectoparasites of neonate Indiana bats, Myotis sodalis (Chiroptera: Vespertilionidae), with description of male of Paraspinturnix globosa (Acari: Spinturnicidae)

A total of 145 ectoparasites was found on five pups of the Indiana bat, Myotis sodalis, from a roost in Warren County, in southwestern Ohio. Most were Macronyssus crosbyi, the common mite of this host, but three males, two nymphs and a nonengorged female of Paraspinturnix globosa were found. Males and nymphs of this species have not previously been reported. In addition, two female adults of Paraspinturnix globosa were found in the anus of one adult female. The male, female deutonymph and nongravid engorged female of P. globosa are described for the first time.     

Innervation and gap junction formation in the myometrium of pregnant little brown bats, Myotis lucifugus

Pregnant Myotis lucifugus were captured in mist nets set outside a large maternity colony and, in most cases, were examined 12-15 hours later. Anterior and posterior halves of uteri were pinned to dental wax and either incubated in glyoxylic acid to produce adrenergic nerve fluorescence, or fixed in buffered glutaraldehyde for electron microscopy. Blood was collected for radioimmunoassay of plasma progesterone. We found no evidence of decreased nerve fluorescence even in late pregnancy when plasma progesterone levels were 10-20 times preovulatory values. Ultrastructural examination also showed no evidence of damage to, or destruction of, nerves in the myometrium. However, we did find gap junctions between myometrial muscle cells during the periparturient period in both normal and aborting bats. Gap junctions began to form several days before term, increased in number until parturition, then decreased dramatically within a day or two thereafter. The percentage of muscle cell plasma membrane involved in gap junction formation was closely correlated with plasma progesterone levels, although whether this is causal or coincidental is not certain. These data do not agree with the conclusions drawn from observations in other mammals that a disappearance of adrenergic nerves from the myometrium is associated with the initiation of parturition, or that gap junction formation is associated with changes in nerve function. They do, however, lend further support to the hypothesis that there is neurogenic control of myometrial contractility in M. lucifugus uteri at term.     

Two novel mastreviruses from chickpea (Cicer arietinum) in Australia

Two novel mastreviruses (genus Mastrevirus; family Geminiviridae), with proposed names chickpea chlorosis virus (CpCV) and chickpea redleaf virus, are described from chickpea (Cicer arietinum) from eastern Australia. The viruses have genomes of 2,582 and 2,605 nucleotides, respectively, and share similar features and organisation with typical dicot-infecting mastreviruses. Two distinct strains of CpCV were suggested by phylogenetic analysis. Additionally, a partial mastrevirus Rep sequence from turnip weed (Rapistrum rugosum) indicated the presence of a distinct strain of Tobacco yellow dwarf virus (TYDV). In phylogenetic analyses, isolates of Bean yellow dwarf virus, Chickpea chlorotic dwarf Pakistan virus and Chickpea chlorotic dwarf Sudan virus from southern and northern Africa and south-central and western Asia clustered separately from these three viruses from Australia. An Australian, eastern Asian, or south-eastern Asian origin for the novel mastreviruses and TYDV is discussed.     

Ten novel mutations found in aniridia

Aniridia (AN) is a sight-threatening congenital ocular disorder characterized by iris hypoplasia, corneal pannus, foveal and optic nerve hypoplasia, cataract formation, and glaucoma. In two-thirds of the patients, AN is inherited in an autosomal dominant fashion with almost complete penetrance but variable expression. The remaining cases are sporadic. Aniridia has been shown to be associated with mutations in the PAX6 gene, located on chromosome 11p13, telomeric to the Wilms' tumor predisposition gene (WT1). This paper describes 14 mutations in the PAX6 gene in patients with AN. Among these 14 mutations, 10 have been unpublished until now. They result most probably in haploinsufficiency and consequently in a reduced protein level of functional PAX6 protein. The mutations reported here are scattered all over the gene, including the paired-box, the glycine-rich region, the homeobox, and the proline–serine–threonine (PST)-rich region. Hum Mutat 12:304–313, 1998.© 1998 Wiley-Liss, Inc     

Two novel uroporphyrinogen decarboxylase (URO-D) mutations causing hepatoerythropoietic porphyria (HEP)

Hepatoerythropoietic porphyria (HEP) is a rare form of porphyria in humans. The disorder is caused by homozygosity or compound heterozygosity for mutations of the uroporphyrinogen decarboxylase (URO-D) gene. Subnormal URO-D activity results in accumulation of uroporphyrin in the liver, which ultimately mediates the photosensitivity that clinically characterizes HEP. Two previously undescribed URO-D mutations found in a 2-year-old Caucasian boy with HEP, a maternal nonsense mutation (Gln71Stop), and a paternal missense mutation (Gly168Arg) are reported here. Recombinant Gly168Arg URO-D retained 65% of wild-type URO-D activity and studies in Epstein-Barr Virus (EBV)-transformed lymphoblasts indicated that protein levels are reduced, suggesting that the mutant protein might be subjected to accelerated turnover. The crystal structure of Gly168Arg was determined both as the apo-enzyme and with the reaction product bound. These studies revealed little distortion of the active site, but a loop containing residues 167-172 was displaced, possibly indicating small changes in the catalytic geometry or in substrate binding or increased accessibility to a cellular proteolytic pathway. A second pregnancy occurred in this family, and in utero genotyping revealed a fetus heterozygous for the maternal nonsense mutation (URO-D genotype WT/Gln71Stop). A healthy infant was born with no clinical evidence of porphyria.     

Ectoparasitic mites (Acari) of sympatric Brazilian free-tailed bats and big brown bats in Alabama.

Seven species of mites were recovered from 133 Brazilian free-tailed bats, Tadarida brasiliensis, and 94 big brown bats, Eptesicus fuscus, from February through November 1990 in colonies that shared roosting space in east-central Alabama. The macronyssid Chiroptonyssus robustipes (Ewing) was the most common mite on T. brasiliensis (964 mites, 87% of bats infested) and on E. fuscus (109 mites, 29% of bats infested). However, C. rubustipes normally is a specific parasite of T. brasiliensis. The macronyssids Steatonyssus ceratognathus (Ewing) and S. occidentalis (Ewing) were recovered from both species of bats in low numbers. S. ceratognathus is not a typical parasite of either species of bat, but S. occidentalis normally is specific to E. fuscus. Predictably, S. occidentalis was most frequently collected from E. fuscus (16 mites, 9% of bats infested), but two specimens were recovered from T. brasiliensis. Five specimens of the laelapid Androlaelaps casalis (Berlese) (a mite that is frequently associated with rodents) and one specimen of the myobiid mite Ewingana (Doreyana) longa (Ewing) (a specific ectoparasite of T. brasiliensis) were also recovered from T. brasiliensis. Singletons of the rosensteiniids Mydopholeus sp. and Nycteriglyphites pennsylvanicus Fain, Lukoschus & Whitaker were the only additional mites collected from E. fuscus; both of these mites have previously been collected from bats or their guano but are recorded here from Alabama for the first time. With respect to ectoparasite cross-infestations, E. fuscus appears to be at greater risk from sharing roots with T. brasiliensis. This is highlighted by the comparatively large numbers of C. robustipes that occurred on E. fuscus and the low numbers of S. occidentalis on T. brasiliensis. Although mites were the only arthropods recovered from bats in this study, a separate survey in 1991 revealed that the bat bug Cimex adjunctus Barber infested some other colonies of T. brasiliensis and E. fuscus in Alabama.     

A novel HLA-DR12 allele (DRB1*1206) found in a Korean B-cell line

At least 6 HLA-DRB1*12 alleles have been identified to date with nucleotide polymorphism occurring at codons 37, 57-58, 60, 67, 85 and 87. In this report, we describe the identification of another new HLA-DRB1*12 allele: DRB1*1206. This novel allele was found in an Epstein-Barr virus (EBV)-transformed Korean B-cell line "K-KT" having the HLA-phenotype A3, 24; B44, 61; Cw3; Bw4, 6; DR12, 13 during full-length cDNA isolation for cell line characterization and for production of HLA-DR recombinant proteins. The allele was identified initially by cycle sequencing of subcloned HLA-DRB full-length cDNA.     

Tick (Acari) infestations of bats in New Mexico

A total of 278 bats belonging to 16 species was examined for ticks from various sites in New Mexico from 1994 to 1998. Seven species of bats were parasitized by ticks: larvae of Ornithodoros kelleyi Cooley & Kohls, Ornithodoros rossi Kohls, Sonenshine & Clifford (Argasidae), or both. Both species of ticks are reported from New Mexico for the first time. Infestation prevalences for parasitized bats ranged from 2 to 25% on different host species for O. kelleyi and from 7 to 25% for O. rossi. The pallid bat, Antrozous pallidus, and the big brown bat. Eptesicus fuscus, were parasitized by both tick species. No distinct host specificity was noted for either tick species.     

Terminal nerve in the mouse-eared bat (Myotis myotis): ontogenetic aspects

As in other mammals, ontogenesis of the terminal nerve (TN) in the mouse-eared bat (Myotis myotis) starts shortly after the formation of the olfactory placode, a derivative of the ectoderm. During development of the olfactory pit, proliferating neuroblasts thicken the placodal epithelium and one cell population migrates toward the rostroventral tip of the telencephalon. Here they accumulate in a primordial terminal ganglion, which successively divides into smaller units. Initial fibers of the TN can be distinguished from olfactory fibers in the mid-embryonic period. The main TN fiber bundle (mfb) originates from the anteriormost ganglion in the nasal roof, whereas one or more inconstant smaller fiber bundles (sfb) originate from one or more smaller ganglia in the basal part of the rostral nasal septum. The fibers of the mfb and sfbs join in the posterior quarter of the nasal roof before reaching the central ganglion (M) located in the meninges medial to the olfactory bulb. From the mid-fetal period onward, a thin TN fiber bundle with some intermingled perikarya connects M to the brain by penetrating its wall rostral to the olfactory tubercle. Additional smaller ganglia may occur in this region. The TN and its ganglia persist in postnatal and adult bats but the number of perikarya is reduced here. Moreover, the different potential functions of the TN are discussed briefly.     

Two novel mutations in a purine nucleoside phosphorylase (PNP)-deficient patient

Purine nucleoside phosphorylase (PNP) deficiency is a rare autosomal recessive disease, which presents clinically as severe combined immunodeficiency (SCID). We report here two novel mutations in the PNP gene that result in SCID phenotype, in a single patient. The maternal-derived allele carries a C to T transition in exon 2 resulting in a premature stop codon at amino acid 57. The paternal-derived mutation is a G to A transition at position +1 in intron 3, causing a complete skipping of exon 3 and a reading frameshift at the exon 2-exon 4 junction. The predicted polypeptide encoded by the aberrantly spliced mRNA terminates prematurely after only 89 amino acids. Both mutations predict severely truncated proteins resulting in a complete deficiency of PNP enzymatic activity, yet the development of profound immunodeficiency in this patient is greatly delayed.