PEG模拟干旱胁迫下8种绿肥作物萌发特性与抗旱性评价

采用不同浓度PEG-6000溶液模拟干旱胁迫的方法,对大麦、黑麦草、二月兰等8种绿肥种子的萌发特性和抗旱性进行了研究。研究结果表明:1)低浓度的PEG-6000对大麦、冬牧70黑麦、毛叶苕子等7种绿肥作物种子的萌发有一定促进作用。2)低浓度的PEG-6000对8种绿肥作物种子胚根的伸长均有一定的促进作用,但对胚芽反而表现出抑制作用。3)随着胁迫程度的加剧,8种绿肥作物种子的发芽率、胚根长、胚芽长、萌发胁迫指数、萌发抗旱指数不断下降,而胚根/胚芽比值却持续上升。4)利用隶属函数分析得出8种绿肥作物种子萌发期的抗旱性强弱次序为:籽粒苋＞白芥末＞黑麦草＞冬牧70黑麦＞沙打旺＞毛叶苕子＞大麦＞二月兰,据其综合评价值分为3类,籽粒苋、白芥末为强抗旱作物种类,黑麦草、冬牧70黑麦、沙打旺、毛叶苕子、大麦为中等抗旱作物种类,二月兰为弱抗旱作物。因此,抗旱性强的籽粒苋、白芥末适宜在黄淮海地区干旱少雨的地区种植。     

Species identification,distribution and abundance of Gerreidae (Teleostei) Bleeker,1859 in the estuaries of Natal

Comparison of published meristic and taxonomic data with measurements taken from specimens collected from Natal estuaries have provided information for an identification key to the Gerres of southern Africa. Body markings were different on each species of fry (< 40 mm S.L.) as well as immatures and adults (> 40 mm S.L.). A comparison of otoliths from the different species showed that they can be used to Identify specimens from a standard length of 20 mm. Turbidity may be a factor limiting the distribution of the genus in estuaries. The known distribution of Gerres species in the estuaries of southern Africa is given. The species of Gerres which is dominant in a particular locality depends on the salinity.     

Frequency of infection by Neospora caninum in wild rodents associated with dairy farms in Aguascalientes,Mexico

The purpose of this study was to detect the presence of Neospora caninum in tissues from wild rodents associated with dairy farms in Aguascalientes, Mexico. Individuals from Mus musculus, Rattus novergicus and Spermophilus variegatus were caught in dairy farms with history of positive status to N. caninum. A total of 33 rodents were caught from which samples were taken of brain, spinal cord, liver and heart and were tested with nested PCR and Immunohistochemistry (IHC). Results obtained with PCR showed that M. musculus had 77% (10/13) positive samples to N. caninum, S. variegatus had 71% (10/14) and R. novergicus had 50% (3/6). The results obtained with IHC showed that 15% (2/13) of M. musculus individuals were positive, while 43% (6/14) of S. variegatus and 33% (2/6) R. novergicus were also positive. This is the first report of the presence of N. caninum in S. variegatus.     

Dynamics of the free-living stages of sheep intestinal parasites on pasture in the North Island of New Zealand. 1. Patterns of seasonal development

Abstract

AIM: To describe the seasonal pattern of development of third-stage infective larvae (L3) from eggs of Teladorsagia (=Ostertagia) circumcincta, Trichostrongylus colubriformis and Haemonchus contortus on pasture in the North Island of New Zealand.

METHODS: Sheep faeces containing known numbers of eggs of all three nematode species were deposited on, or buried in, pasture plots at three sites, viz coastal Manawatu, Upper Hutt Valley, and East Cape hill country. Development was measured by recovering L3 from faeces, herbage and soil 28–31 days after deposition on 13–18 occasions, between January 2005 and July 2006. Analysis of the number of larvae recovered used a mixed model including number of eggs deposited, weight of faeces recovered (an assumed indicator of earthworm activity), site, contamination date, and position of deposited faeces, i.e. on the surface or buried.

RESULTS: There was a significant effect of contamination date on development of all three species, with maximum numbers ofL3 developing between late spring (November) and early autumn (March), and minimum numbers in June and July. There were large differences between species, with H. contortus exhibiting a long period (April to October) where development was close to zero, whereas T. circumcincta developed to some extent all year round. Development of T. colubriformis was intermediate between the other two species.

Burying faeces containing nematode eggs increased the number of L3 recovered compared with surface deposition (p≤0.001), although there were a small number of exceptions involving only T. colubriformis. The weight of faeces recovered at harvest, which was assumed to be an indication of earthworm activity, was correlated with the number of L3 recovered for all species (p<0.001). In a separate analysis, earthworms were assumed tohave been active if <5 g faeces remained at harvest. Where this occurred, the number of L3 of T. colubriformis and T.circumcincta recovered was reduced by 56% and 58%, respectively (p<0.001).

CONCLUSIONS: A marked seasonal pattern of development was observed for all three species, with the most larvae developing in spring-early autumn and the least in winter. This seasonal pattern was most pronounced in H. contortus and least obvious in T. circumcincta. Burying faeces containing eggs generally resulted in more L3 being recovered, whilst the apparent activity of earthworms resulted in fewer larvae being recovered.     

Dynamics of the free-living stages of sheep intestinal parasites on pasture in the North Island of New Zealand. 2. Weather variables associated with development

Abstract

AIM: To identify weather variables associated with the development of eggs of Teladorsagia (=Ostertagia) circumcincta, Trichostrongylus colubriformis and Haemonchus contortus to third-stage infective larvae (L3) under a range of climatic conditions on pasture in the North Island of New Zealand.

METHODS: Sheep faeces containing known numbers of eggs of all three nematode species were deposited on, or buried in, pasture plots at three sites, viz coastal Manawatu, Upper Hutt Valley, and East Cape hill country. Development was measured by recovering L3 from faeces, herbage and soil 28–31 days after deposition, on 13–18 occasions between January 2005 until July 2006. Weather data were recorded at each site, and the association between weather variables and number of L3 recovered was analysed using subsets regression to select best-fitting models from several candidate variables, after adjustment for efficiency of recovery of L3. A multiple linear regression model was then developed for each species, to select weather variables that had both significant and substantive effects on the number of L3 recovered.

RESULTS: For all species, mean daily temperature was the bestpredictor of the number of L3 recovered (p=0.001). For T.circumcincta the final model included mean daily temperature and soil temperature (R²=51%), and for T. colubriformis the model only included mean daily temperature (R²=55%). For development of H. contortus, mean daily temperature was the most significant variable, but moisture in the form of rainfall entropy, i.e. the temporal distribution of rainfall, overthe first 14 days was also significant in the final model (R²=34%).

CONCLUSIONS: Temperature was the most important determinant of developmental success of free-living nematodes on pasture at the study sites, and probably also for other parts ofNew Zealand with similar climates. Moisture was not significant in the development of T. circumcincta or T.colubriformis, implying that under the generally moist temperate climate in New Zealand, moisture is seldom limiting for development of these species. Haemonchus contortusappeared to be sensitive to moisture availability and precipitation in the first 14 days after deposition of faeces. The results of this study will be further developed as part of species-specific climate-driven models of parasite development in New Zealand.     

DNA methylation and expression of imprinted genes are associated with the viability of different sexual cloned buffaloes

The DNA methylation of imprinted genes is an important way to regulate epigenetic reprogramming of donor cells in somatic cell nuclear transfer (SCNT). However, the effects of sexual distinction on the DNA methylation of imprinted genes in cloned animals have seldom been reported. In this study, we analysed the DNA methylation status of three imprinted genes (Xist, IGF2 and H19) from liveborn cloned buffaloes (L group, three female and three male), stillborn cloned buffaloes (S group, three female and three male) and natural reproduction buffaloes (N group, three female and three male), using bisulphite sequencing polymerase chain reaction (BS‐PCR). The expression levels of these imprinted genes were also investigated by quantitative real‐time PCR (QRT‐PCR). The DNA methylation levels of H19 were not significantly different among the groups. However, the Xist in female and IGF2 in male of the S group were found to be significantly hypomethylated in comparison with the same sexual buffaloes in L group and N group (p < .05). Furthermore, the expression levels of Xist, IGF2 and H19 in the stillborn female cloned buffaloes of S group were significantly higher than that of the female buffaloes in the L group and N group (p < .05). The expression levels of IGF2 and H19 in the stillborn male cloned buffaloes in the S group were significantly higher than that of the male buffaloes in the L group and N group (p < .05). These results indicate that Xist may be associated with the viability of female cloned buffaloes, and IGF2 may also be related to the viability of male cloned buffaloes.     

Sessile peritrichs (Ciliophora: Peritricha) from freshwater fish in the Transvaal,South Africa

Sessile peritrichs (Ciliophora: Peritricha) were collected from freshwater fish in lakes, rivers, streams and fish-ponds in the Transvaal, South Africa Nine species of the genus Apiosoma Blanchard, 1885 are described, i.e. seven new species; A. caulata sp.n., A. curvinucleata sp.n, A. micralesti sp.n., A. mothlapitsis sp.n., A. obliqua sp.n., A. phiala sp.n. and A. viridis sp.n., and two known species; A. nasal is (Timofeev, 1962) and A. piscícola Blanchard, 1885. One new species of the genus Ambiphrya Baabe, 1952 is described, i.e. A. neobolae sp.n. A new sgenus, Scopulata gen.n. is proposed for species with a broad scopula and compact macronucleus. Two previously described Scyphidia species are incorporated in this genus, i.e. S. dermata (Viljoen & Van As, 1983) comb.n. and S. epibranchialis (Viljoen & Van As, 1983) comb.n., as well as a new species, S. constricta sp.n. Compendiums of all the known fish-associated species of these genera are provided.     

Freshwater nematodes from South Africa. 8. New and known species of Actinolaimoidea

Afractinolaimus Andrássy, 1970 is regarded as valid, its diagnosis slightly emended, and the following species transferred to it: Paractlnolaimus capensis, P. minor, P. zairensis and P. magaliesmontanus. The name of the type species thus becomes A. magaliesmontanus, since it has priority and is regarded as conspecific with A. noblei, the designated type. Additional biometrical data and distribution records are presented for A. magaliesmontanus. A. zairensis is reported from South Africa. Information is given for several populations of P. microdentatus. Two new species of Neoactinolaimus are described: N. brachydorus n. sp. is characterized by an exceptionally short and stout odontostyle, and N. barbieri n. sp. is distinguished from the closely related N. vaalensis by a thicker cuticle, more prominently set-off head, corrugated vestibule, longer oesophagus and several other minor morphological differences. Egtitus Thorne, 1967 is regarded as valid, and the following combination proposed: Egtitus zealandicus (Clark, 1963) n. comb, (synonym: Actinolaimus zealandicus).     

Distribution of Myxobolus cerebralis in Salmonid Fishes in Montana

Abstract

Over an approximately 2-year period, 20,974 fish (trout and other salmonid species) from 230 separate waters (creeks, rivers, lakes, reservoirs, ponds, hatcheries, and irrigation ditches) within 21 of the 22 major drainages in Montana were examined for Myxobolus cerebralis. Nine of the major river drainages have waters containing infected fish: Beaverhead, Big Hole, Blackfoot, Clark Fork above the Bitterroot River, Flathead below the south fork of the Flathead River, Jefferson, Madison, Missouri above the Marias River, and Yellowstone above the Bighorn River. The Beaverhead, Clark Fork above the Bitterroot River, Jefferson, Madison, and Missouri above the Marias River have the greatest number of waters containing fish infected with M. cerebralis. Comparisons of infection levels (number of pooled samples that contain fish infected with M. cerebralis) between species among these drainages show significantly lower levels of infection in brown trout Salmo trutta and rainbow trout Oncorhynchus mykiss in the Missouri above the Marias River drainage and significantly higher levels of infection of rainbow trout in the Jefferson. Comparisons of differences in infection levels between drainages among species show that, in the Beaverhead, Clark Fork above the Bitterroot River, and Madison, infection levels in brown trout are significantly higher than in rainbow trout. This is partially attributed to losses of juvenile rainbow trout because of M. cerebralis infection, leading to biased samples. Histopathologic studies showed lesions were consistently less severe in brown trout than other species and occurred in a different location (gill arches versus ventral calvarium). In six of the nine affected drainages (Beaverhead, Blackfoot, Clark Fork above the Bitterroot River, Flathead below the South Fork, Jefferson, and Madison), infected fish were found at or near the time that intensive sampling was initiated in the spring of 1995. In the three remaining affected drainages (Missouri above the Marias River, Yellowstone above the Bighorn River, and the Big Hole), infected fish were not identified until at least 15 months after the initiation of widespread testing. This indicates that in the first six drainages listed above, the infection was well established prior to 1995 but spread into the last three drainages in the ensuing months. Methods of transmission and the sources of infection are unknown, although the absence of infected fish in state, private, and federal hatcheries in Montana indicates hatchery fish from these sources are not likely to be responsible.     

Occurrence and characterization of gastric Helicobacter spp. in naturally infected dogs

Helicobacter-like organisms are frequently observed in the stomach of dogs but the relationship between these microorganisms and gastric pathology has not been clearly established. Different species of helicobacters are known to be present in the canine stomach but their specific prevalence in naturally infected dogs is unknown. The aims of this study were to isolate and characterize helicobacters in canine gastric biopsies, to compare the commonly used tests for the identification of Helicobacter spp. and to determine the occurrence of these species in dogs. Twenty-three out of 25 dogs (92%) were positive for Helicobacter-like organisms in cytological screening. Culture was successful from biopsies of 5/25 dogs. The isolates were analyzed by electron microscopy, biochemical and physiological tests, whole protein analysis and 16S rDNA sequencing. Helicobacter felis was identified in four samples and Helicobacter bizzozeronii in one sample. Only the whole protein analysis in combination with electron microscopy was able to clearly discriminate the two species. Compared to the high prevalence of Helicobacter-like organisms, the occurrence of H. felis and H. bizzozeronii, was low (17 and 4%, respectively). No Flexispira rappini-like organisms or H. salomonis were detected. Electron microscopy revealed that H. bizzozeronii-like microorganisms were present in three additional biopsies where we were unable to culture any Helicobacter-like organisms. These observations indicate that in the stomach of dogs not all helicobacters are culturable. The unculturable bacteria appeared to be the prevalent ones and may represent different spiral organisms. The presence of distinct helicobacters with different characteristics can reflect different roles in the pathogenesis of canine gastric disease.     

Validation of the periodicity of growth zone formation in the otoliths of four fish species from the Upper Zambezi ecoregion,southern Africa

In the Upper Zambezi and Okavango ecoregions, Brycinus lateralis, Hepsetus cuvieri, Schilbe intermedius and Serranochromis macrocephalus are important in subsistence fisheries, while S. intermedius and S. macrocephalus are often caught in commercial catches. Despite their importance, there is little information on their age and growth and age validation. Growth zone deposition rate in otoliths of B. lateralis, H. cuvieri, S. intermedius and S. macrocephalus was validated as annual using edge analysis in this study. Annual deposition of growth zones was corroborated by a fluorochrome marking experiment. Both methods demonstrated that one growth zone was deposited annually. Edge analysis demonstrated that the highest proportion of opaque zones was detected between August and December, most likely as a result of slow growth during lower temperature water during the preceding winter. With growth zone deposition validated as annual, future research should focus on determining the growth, maturity and mortality rates of these species, parameters which are important for their management.     

Has Sarcocystis neurona Dubey et al., 1991 (Sporozoa: Apicomplexa: Sarcocystidae) cospeciated with its intermediate hosts?

The question of how Sarcocystis neurona is able to overcome species barrier and adapt to new hosts is central to the understanding of both the evolutionary origin of S. neurona and the prediction of its field host range. Therefore, it is worth reviewing current knowledge on S. neurona host specificity. The available host range data for S. neurona are discussed in relation to a subject of evolutionary importance—specialist or generalist and its implications to understand the strategies of host adaptation. Current evidences demonstrate that a wide range of hosts exists for S. neurona. This parasite tends to be highly specific for its definitive host but much less so for its intermediate host (I.H.). The unique specificity of S. neurona for its definitive host may be mediated by a probable long coevolutionary relationship of the parasite and carnivores in a restricted ecological niche ‘New World’. This might be taken as evidence that carnivores are the ‘original’ host group for S. neurona. Rather, the capacity of S. neurona to exploit an unusually large number of I.H. species probably indicates that S. neurona maintains non-specificity to its I.H. as an adaptive response to insure the survival of the parasite in areas in which the ‘preferred’ host is not available. This review concludes with the view that adaptation of S. neurona to a new host is a complex interplay that involves a large number of determinants.     

XX/XY mosaicism in lymphocyte cultures from a pig with freemartin characteristics

Sir. — Cultural confirmation of venereal campylobacteriosis in cattle can only be achieved if samples arc cultured on the day of collection or a suitable transport medium is used. Clark et al.⁽³⁾ have developed a selective enrichment medium for Campylobacter fetus venerealis which is also an effective transport medium ⁽⁸⁾. However, this transport medium is very cumbersome to prepare in that it requires a special gas mixture and large quantities of fresh bovine serum. This letter reports on the evaluation of Clark’s ⁽⁸⁾, cooked meat medium with antibiotics (CMMA), Weybridge ⁽⁴⁾, modified EMJH ⁽⁵⁾, Stuart’s (Oxford) and Cairy Blair (Oxoid) media, as transport media for C. fetus veneralis.     

Morphologic,morphometric and contour shape variations of sagittal otoliths of Lepidorhombus spp. in the Aegean Sea

Otoliths are calcareous anatomical structures in the inner ear of fishes, and they can be used in the discrimination of fish species and stocks due to their species-specific shape. Sagittal otoliths in the Lepidorhombus whiffiagonis and Lepidorhombus boscii, two flatfish species of the same genus distributed in the Aegean Sea, were compared using morphology, morphometry and contour shape analyses. Blind and eyed side otoliths of Lepidorhombus species were evaluated separately due to their morphological features and statistical differences in morphometric measurements (p < 0.05). Four analysis groups were formed: blind side otolith of L. whiffiagonis, eyed side otolith of L. whiffiagonis, blind side otolith of L. boscii and eyed side otolith of L. boscii. Morphometric differences in otolith shape were performed by canonical discriminant analysis, and the first discriminant axis explained 97.4% (Wilks λ = 0.270) of the variance between the groups, and the second axis explained 2.3% (Wilks λ = 0.620). Classification success between Lepidorhombus species based on CDA is 100%, and the overall CDA classification score between groups is 76.5%. The wavelet functions obtained in the contour analysis showed high variability in the anterior, posterior-dorsal, antero-dorsal and ventral regions of the otoliths among these four analysis groups. Consequently, it is thought that this study will contribute to the taxonomic classification of fish with the morphological and morphometric differences in the blind and eyed side otoliths and the analyses made with the high discrimination success obtained in Lepidorhombus species.     

A new euryhaline copepod from Cape Town: Halicyclops dedeckeri n.sp. (Copepoda:Cyclopoida)

A new species of the widely distributed cyclopid genus Halicyclops Norman is described from hypersaline summer pools on the bed of Milnerton Lagoon, Cape Town, South Africa. H. dedeckeri n. sp. appears to be most closely related to H. thermophilus Kiefer, 1929 and H. spinifer Kiefer, 1935 in that the females of all three species bear a sharp chitinous spine on each side of the genital double-somite. Unlike H. thermophilus and H. spinifer, the new species bears one spiniform and one normal seta (in addition to the three spines) on the P4 enp 3 in both sexes. The new species is being mass-cultured in laboratory tanks for use in feeding studies on marine fish larvae.     

Alternate Pathway of Infection with Hepatozoon americanum and the Epidemiologic Importance of Predation

Background: The range of American canine hepatozoonosis (ACH) is expanding from the southern USA northward. Transmission of Hepatozoon americanum occurs by ingestion of infected Gulf Coast ticks, Amblyomma maculatum. The source of the protozoan for the tick remains undetermined; infected dogs are unusual hosts for the tick. Objective: Compare possible sources of infection by field investigations of 2 multiple‐dog outbreaks of ACH. Animals: Twenty‐eight privately owned dogs (Canis familiaris), 1 coyote (Canis latrans), 31 wild‐trapped cotton rats (Sigmodon hispidus), 24 wild‐trapped field mice (Peromyscus leucopus), and 9 wild‐caught rabbits (Sylvilagus spp.) from sites in eastern Oklahoma were monitored for hepatozoonosis. Six laboratory‐raised cotton rats (S. hispidus), 6 Sprague‐Dawley rats (Rattus norvegicus), 6 C57BL/6J‐Lystbg‐J/J mice (Mus musculus), 6 outbred white mice (M. musculus), 6 New Zealand white rabbits (Oryctolagus cuniculus), and 2 dogs were acquired through commercial vendors for experimental transmission trials of H. americanum. Methods: Four of 15 dogs in a rural neighborhood and 5/12 hunting Beagles were confirmed to be infected by blood smear examination, muscle biopsy, and polymerase chain reaction assay of the 18S rRNA gene of Hepatozoon species. Histories and tick host preferences led to field collections of common prey of canids and experimental transmission trials of H. americanum to selected prey (M. musculus, S. hispidus, R. norvegicus, and O. cuniculus). Results: Dogs with ready access to prey (4/15 dogs) or that were fed prey retrieved from hunts (5/12 hunting Beagles) became infected, providing evidence that predation is an important epidemiologic component of ACH infection. Experimental transmission studies identified a quiescent, infectious stage (cystozoite) of the parasite that provides an alternate mode of transmission to canids through predation, demonstrating that cotton rats, mice, and rabbits but not brown rats may act as paratenic hosts of H. americanum. Conclusions and Clinical Importance: Predation of prey harboring infected A. maculatum or containing cystozoites of H. americanum in their tissues provide 2 modes of transmission of ACH to dogs, putting unconfined dogs at increased risk of infection in endemic areas.     

The Breeding Season of the Multimammate Mouse Praomys (Mastomys) Natalensis (A. Smith) in the Transvaal Highveld

Certain aquatic Oligochaeta live successfully in organic-polluted waters while others cannot tolerate such conditions. Tubifex tempUtoni Southern and Llmnodrttus hoffmeisteri (Clap.), two non-gilled species, are compared to Branchiura sowerbyi (Beddard) and Dero nivea (Aiyer), two gilled species, concerning their haemoglobin concentration, the influence of low oxygen availability on the respiration rate and their reactions in an oxygen gradient. In contrast with L. hoffmeisteri and T. templetoni where a very narrow range of respiration dependency (pO₂, 0-12 mm Hg) is found, B. sowerbyi and D. nivea have a level of respiratory dependence from pO₂, 0-131 mm Hg. Haemoglobin concentrations are 6,238 - and 6,452 g/100 cm² blood in L. hoffmeisteri and T. templetoni and 3,011 and 1,611 g/100 cm² in B. sowerbyi and D. nivea respectively. In the oxygen gradient all four species have a preference for water with a high oxygen content.     

Identification of Streptococcus dysgalactiae Strains of Lancefield's group C,G and L by Polymerase Chain Reaction

Streptococcus dysgalactiae serogroup C, G and L strains were investigated by polymerase chain reaction (PCR) using oligonucleotide primers designed according to species‐specific parts of the 16S–23S rDNA intergenic spacer region. The oligonucleotide primers with specificity for the 16S–23S rDNA intergenic spacer region allowed a correct identification of all S. dysgalactiae serogroups C, G and L strains investigated. No cross‐reactivities could be observed with any of the control strains indicating the usefulness of PCR‐technology to identify the serologically heterogeneous species S. dysgalactiae.     

The ecology of Sphyraena barracuda (Osteichthyes: Perciformes) in the Kosi system with notes on the Sphyraenidae of other Natal estuaries

The ecology of Sphyraena barracuda was studied in the Kosi estuary system of Natal. Fry enter from the sea at a length of 20 mm and live in shallow marginal weed-beds until a length of 80 mm when they move to fringing Phragmites reed-beds. After reaching 300 mm they move to open water areas. No specimens longer than 500 mm were recorded and none were mature. Growth is rapid, they attain 180 mm in the first six months and after two years measure 400-500 mm. They leave the estuary after about two years. Their significance in the estuarine fish community is due to their predation, at all sizes, on a variety of estuarine-dependent juvenile fishes (e.g. Rhabdosargus spp., Mugilidae) and adults of estuarine species (e.g. Ambassis spp.). S. barracuda was recorded at salinities between 4°/oo and 35°/oo, only in very clear water and at temperatures from 14,4 °C to 37 °C. Sphyraena bleekeri and S. qenie, which are also clear-water species, were found at Kosi in salinities from 0,5 to 35%o. They are also piscivorous on a range of small teleosts. Sphyraena iello is the most widespread barracuda in Natal estuaries, but only juveniles were recorded. This species is tolerant of a wide range of turbidities and salinities which enables it to live under most estuarine conditions. The distribution, ecology and interspecific interactions among Sphyraenidae in Natal estuaries are discussed in relation to their feeding biology, habitat preferenda and physical tolerances.